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Background: The increasing problem of drug resistance to most of the available antimalarial therapies is a recurring issue. To overcome this issue, new antiplasmodial drugs are needed. Many plants have been use for treatment of malaria by traditional healer in India. The purpose of this research was to evaluate seven medicinal plants used against malaria by indigenous people in India, validate their use and identify the plant with the best antiplasmodial properties. Methods: Methanol, hydro alcoholic (60:40) and aqueous extracts from seven ethnomedicinal plants were screened on chloroquine-sensitive (MRC-2) and chloroquine-resistant (RKL-9) strain of Plasmodium falciparum by schizont maturation inhibition assay and inhibition of Plasmodium falciparum lactate dehydrogenase (PfLDH) activity. Results: Out of 21 extracts, hydro alcohol extract of B. variegate showed the highest inhibition of schizont maturation with 81.77±0.96% against MRC-2 and 78.08±3.76% against RKL-9 strain. In PfLDH inhibition assay, Methanol and hydro alcohol extract of B. variegate showed moderate antiplasmodial activity with IC50 24.65 μg/mL and 39.42 μg/mL, respectively against MRC-2 strain. Methanol extract of B. variegate also exhibited moderate activity with IC50 28.71 μg/ml against RKL-9 strain. Conclusion: The present study provided scientific evidence for indigenous Indian medicinal plants as a source of antiplasmodial agents. This finding supports the continued exploration of traditional medicines in the search for new antimalarial agents. The compounds responsible for the observed antiplasmodial effects in B. variegate are under investigation.
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Page 1/11
Antiplasmodial Potential of Traditional Phytotherapy
of Some Remedies Used in The Treatment of
Malaria in India
Navin Sheth ( jmvadalia15@gmail.com )
Gujarat Technological University
Jigna Vadalia
Department of Pharmaceutical Sciences, Saurashtra University, Rajkot, Gujarat
Jalpa Sanandia
Department of Pharmaceutical Sciences, Saurashtra University
Vaibhav Bhatt
Department of Pharmaceutical Sciences, Saurashtra University
Research
Keywords: antimalarial activity, Plasmodium falciparum, schizont maturation inhibition assay, Parasite
lactate dehydrogenase (PfLDH) assay, Indian plants
DOI: https://doi.org/10.21203/rs.3.rs-66865/v1
License: This work is licensed under a Creative Commons Attribution 4.0 International License.
Read Full License
Page 2/11
Abstract
Background: The increasing problem of drug resistance to most of the available antimalarial therapies is
a recurring issue. To overcome this issue, new antiplasmodial drugs are needed. Many plants have been
use for treatment of malaria by traditional healer in India. The purpose of this research was to evaluate
seven medicinal plants used against malaria by indigenous people in India, validate their use and identify
the plant with the best antiplasmodial properties.
Methods: Methanol, hydro alcoholic (60:40) and aqueous extracts from seven ethnomedicinal plants
were screened on chloroquine-sensitive (MRC-2) and chloroquine-resistant (RKL-9) strain of
Plasmodium
falciparum
by schizont maturation inhibition assay and inhibition of
Plasmodium falciparum
lactate
dehydrogenase (
Pf
LDH) activity.
Results: Out of 21 extracts, hydro alcohol extract of
B. variegate
showed the highest inhibition of schizont
maturation with 81.77±0.96% against MRC-2 and 78.08±3.76% against RKL-9 strain. In
Pf
LDH inhibition
assay, Methanol and hydro alcohol extract of
B. variegate
showed moderate antiplasmodial activity with
IC50 24.65 μ g/mL and 39.42 μg/mL, respectively against MRC-2 strain. Methanol extract of
B. variegate
also exhibited moderate activity with IC50 28.71 μg/ml against RKL-9 strain.
Conclusion: The present study provided scientic evidence for indigenous Indian medicinal plants as a
source of antiplasmodial agents. This nding supports the continued exploration of traditional medicines
in the search for new antimalarial agents. The compounds responsible for the observed antiplasmodial
effects in
B. variegate
are under investigation.
Background
Malaria still poses a threat to the health of residents and travellers in tropical and subtropical countries.
There were 405 000 malaria deaths recorded globally in 2018 as per World Health Organization. Children
aged below 5years were more affected, as they accounted for 67 percent of malaria death globally in
2018. The African Region accounted for the highest, 94 percent of malaria deaths in 2018 [1]. Two of the
main reasons for malaria perseverance are increasing
P. falciparum
drug resistance and insecticide
resistance of mosquito vectors [2]. Amongst the ve existing species of Plasmodium causing malaria in
humans,
P. falciparum
is the most virulent and widespread infectious due to the resistance of the parasite
to most of the available antimalarial drugs [3]. The development of resistance to golden therapeutic
drugs like chloroquine and controlled use of new artemisinin analogs have created an urgent need to
discover new antimalarial agents [4]. Artemisinin resistance has been reported rst from clinical cases on
the Cambodia–Thailand border and then spread to other countries [5]. The need for new drugs, preferably
with new modes of action is therefore needed [6]. Plant species are the best source for chemotherapeutic
prophylactics for malaria treatment [7]. Two most available medicine quinine and artemisinin invented
from the scientic study of traditional remedies of various cultures. Many plant species are reported for
Page 3/11
the treatment of malaria and fever [8]. Plants have been proven for their value as a rich source of a
therapeutic molecule and many currently available drugs are natural products-derived compounds [9].
In this present study, the selection of plants to be screened for antimalarial activity was done based on
the traditional claim of particular plants for the treatment of malaria. Here, we studied plants mentioned
either in the traditional text or used by traditional healers of India for ''
vishamjwara
(malaria)'' and
evaluated them
in vitro
for antimalarial activity.
Methods
Plant materials and chemicals
The plants were collected in the month of October-November from Saurashtra University Campus and the
surrounding area, Rajkot. The parts of the plant used, their family, local name and traditional uses of the
seven selected plants are stated in Table 1. The plant species were identied by Dr. Vrunda Thakar,
Department of Bioscience, Saurashtra University, Rajkot. Voucher specimens were deposited to the
Department of Pharmaceutical Sciences, Saurashtra University, Rajkot. The plant materials were dried
under shade, stored in an airtight container at room temperature and powdered to 60 meshes when
required.
RPMI-1640 media with phenol red, glutamine and HEPES buffer, albumax II, hypoxanthine, Sodium
bicarbonate (NAHCO3), sorbitol, Dimethyl sulphoxide (DMSO) and Giemsa stain were purchased from
Himedia Laboratories. Quinine diphosphate and artemisinin were purchased from Sigma-Aldrich. All other
chemicals were of analytical grade.
Preparation of crude plant extracts
25 gm dry powder of each plant species were extracted with methanol, methanol: water (60:40) and water
by hot maceration at 60°C in a water bath for 6 h followed by 48 h maceration with continuous shaking in
a rotary shaker at 100 RPM. This process was repeated three times with the same solvent. The extracts
were ltered, combined and the solvent is removed using a rotary evaporator under vacuum. The dried
extracts were stored at 4°C.
In vitro
cultivation of
Plasmodium falciparum
Chloroquine sensitive (MRC-2) and chloroquine-resistant (RKL-9) strain of
P. falciparum
were procured
from the National Institute of Malaria Research (NIMR), New Delhi, India. Cultures were maintained
in
vitro
by a modied method of Trager and Jenson [20] in human red blood cells 5% hematocrit using
RPMI-1640 with phenol red, glutamine and 20 mM HEPES buffer as a culture medium supplemented with
0.4% albumax II, 0.0025% hypoxanthine, 0.21% NAHCO3 and 40 µg/mL gentamycin. Slides of culture
were observed after 6, 12, 24 and 48 h for regular development of parasite stages. Parasites were
synchronized by repeated treatment of 5% sorbitol.
Page 4/11
Schizont maturation inhibition assay
The antiplasmodial activity of extracts of the plants was assessed by the schizont maturation inhibition
assay [21]. 100 µL of highly synchronized parasite culture with 1-1.5% parasitemia and 5% hematocrit
was added in each well in 96 well plates along with 10µl of different extract of 10 and 30 µg/mL. Quinine
diphosphate and artemisinin were used as positive control. Infected and uninfected erythrocytes were
used as control and negative controls. Each concentration was repeated three times (triplicate each). The
plates were kept for 48 h under 37°C, 5% CO2 atmosphere. After 48 h of incubation, thin blood smears
were prepared from each experimental and control wells on pre-labeled slides and xed in methanol.
Dried slides were stained with Giemsa stain and observe under 100 x under a microscope (LEICA DM1000
LED). Numbers of schizonts were counted per 200 asexual stages of parasites. The inhibition of schizont
development in comparison to the control wells was determined.
Lactate dehydrogenase inhibition assay for
P. falciparum
The antiplasmodial activity was analyzed by measuring Plasmodium lactate dehydrogenase (
Pf
LDH)
activity [22, 23]. 100 µL of highly synchronous ring stage culture with 1% parasitemia and 3% hematocrit
was incubated in 96-well microtitre plate with 10µL of different concentrations of test compounds (5-500
µg/mL) at 37 °C for 48 h. Quinine diphosphate and artemisinin were used as positive control. Infected
and uninfected erythrocytes were used as control and negative controls. After incubation, plates were
subjected to three 30 min freeze-thaw cycles, then 20 μL of supernatant were transferred to another
microtitre plate containing 100 μL of Malstat reagent (0.125% Triton X-100, 130 mM L-lactic acid, 30 mM
Tris buffer and 0.62 μM 3-acetylpyridine adenine dinucleotide; pH 9) and 25 μL of NBT-PES (1.9 μM nitro
blue tetrazolium and 0.24 μM phenazine ethosulphate) solution per well. The plate was incubated in the
dark for 30 min and absorbance was recorded at 630 nm using a microplate reader (BioTek ELx800).
Control parasite cultures devoid of plant extracts or drugs were referred to as 100%
Pf
LDH activity. IC50
values were calculated by the dose-response curve. Each analysis was performed in triplicate (n =3).
Statistical analysis
Results were expressed as means followed by a standard deviation. IC50 was determined from the
resulting dose-response curve non-linear regression using Prism 5 for Windows, Version 5.02 (Graph Pad
Software, Inc) program. . Comparisons were made between disease control and treatments by using one
way analysis of variance (ANOVA) followed by Tukey′s multiple comparison tests., with the level of
signicance set at *
p
≤0.05; **
p
≤0.01; ***
p
≤0.001.
Results
Table 2 shows the herbarium number and the amounts of raw material used for extraction against the
percentage yields of the extracts.
Schizont maturation inhibition
Page 5/11
Twenty-one extracts were prepared from the seven plant species were rst tested for schizont maturation
inhibition assay at two concentrations 10 µg/mL and 30 µg/mL against Chloroquine sensitive (MRC-2)
and chloroquine-resistant (RKL-9) strain of
P. falciparum
(Fig 1.). Parasites of all stages were observed in
the control culture wells. Hydro-alcohol extract (30 µg/mL) of
B. variegate
showed more number of ring
stages and with higher inhibition of schizont maturation with 81.77 ± 0.96% and 78.08 ± 3.76% against
chloroquine-sensitive MRC-2 and Chloroquine-resistant (RKL-9) of
P. falciparum
respectively. The
methanol extract (30 µg/mL) of
B. variegate
showed inhibition of schizont maturation with 76.40 ± 1.69%
and 69.66 ± 2.81% against chloroquine-sensitive MRC-2 and Chloroquine-resistant (RKL-9) of
P.
falciparum
respectively. Hydroalcoholic extract of
A. excelsa
and
F. arabica
also demonstrated interesting
schizont maturation inhibition activity against both the strain. Microscopic examination of uninfected
erythrocytes incubated with the extracts of these plants showed no morphological differences after 48 h
of incubation.
Inhibition of
Pf
LDH
All the extracts showed dose-dependent inhibition of
P. falciparum
by inhibiting
Pf
LDH against both the
strains. As per WHO guideline and some earlier research, [24] activity criteria for assay of plant extracts
were dened as high when IC50<10 µg/ml, an IC50 10-50 µg/mL; moderate activity, low activity when an
IC50 between 50 - 100 μg/mL, and an IC50> 100 μg/mL; lack of activity. Methanol and hydro alcohol
extract of
B. variegate
showed moderate antiplasmodial activity with IC50 24.65 μg/mL and 39.42 μ g/mL
respectively against MRC-2 strain. The methanol extract of
B. variegate
also exhibited moderate activity
with IC50 28.71 μg/mL against RKL-9 strain. The extracts were more active on MRC-2 than RKL-9 strain.
IC50 of reference drug Quinine diphosphate and artemisinin for both the strain were stated in Table 3.
Discussion
Malaria is a highly destructive parasitic disease affecting many developing countries. Drug resistance to
available therapy is a big challenge amongst all scientists. To overcome this resistance problem,
traditional health care systems can be a better source for developing new molecules with more ecacies
and minimum toxicity. Seven plant species were selected from the traditional system of India which has
traditional use against malarial fever. Seven plant species have been tested rst time for both
P.
falciparum
strains. The present study reveals that leaves of
B. variegate
having the best activity amongst
seven screened plants in schizont maturation inhibition assay and
Pf
LDH inhibition assay In the earlier
report, some new secondary metabolites dihydrodibenzoxepins and avanones were isolated from
Bauhinia
species and exhibited antimalarial activity (IC50 5.8–11.2µM) [25]. Phenolic metabolites and
avonoids seem to be the main active principles detected by
Bauhinia
spp. Leaves extract of
B. variegate
exhibited better schizonticidal action against
P. berghei
in mice screened by Peter's 4-day test [26]. The
plant crude extracts screened were less active than reference drugs quinine and artemisinin, may be due
to the composition of many heterogeneous compounds present with active compounds and active
principles might show higher activity in their pure form. The screening results do not reect an evaluation
Page 6/11
of the traditional use of the plants. A majority of the plants selected did not show
in vitro
antiplasmodial
activity, even though a strong correlation with malaria and its treatment in traditional medicine in India.
This may be due to these plants may act as antipyretics or cure the symptoms of malaria, rather than
having activity on malaria parasite. All the water extract tested were inactive, some previous studies
manifested a similar effect of crude water extracts less potent than their corresponding methanolic
extracts. [27, 28]
Conclusion
B. variegate
has good potential and can be used for the development of an antimalarial drug. Preliminary
screening results are suciently interesting for further phytochemical investigations to identify the
chemical compounds responsible for the antimalarial activity and obtain standardized preparations.
Bioassay-guided fractionation of the leaves of
B. variegate
extracts is in progress to isolate and identify
the active compounds.
Declarations
Ethics approval and consent to participate: Not applicable
Consent for publication: Not applicable
Availability of data and materials
The datasets used and analysed during the current study are available from the corresponding author on
reasonable request.
Competing interests Not applicable
Funding Not applicable
Authors' contributions
JV designed, performed the laboratory work, analyzed data and drafted the manuscript. NS conceptualize
as well as supervised the study and revised the manuscript. VB supervised study and revised the
manuscript. JS participated in the laboratory work. All authors read and approved the nal manuscript.
Acknowledgements
The authors acknowledge the constant support of Dr. Mihir Raval, Head of the Department of
Pharmaceutical Sciences, Saurashtra University, Rajkot, India throughout the entire research work.
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Tables
Page 9/11
Table 1
Plants with their families, local name, parts of plant used and traditional uses
Botanical
name Family Local Name Plant
part Traditional use and References
Ailanthus
Excelsa
(AE) Simaroubaceae Arduso Leaves Leaf juice and bark juice use in
forests located near Bhubaneswar,
Orissa, India. [10, 11, 12]
Bahaunia
variegate
(BV)
Caesalpiniaceae Kachnar Leaves South Indians used leaf decoction to
relieve headaches in malarial fever
[13, 14]
Cesealpinia
crista
(CC) Caesalpiniaceae Fever nut Leaves In Assam, India, the leaves and seeds
are traditionally used to treat malarial
fever [10, 15]
Alstonia
scholaris
(AS)
Apocynaceae Saptaparni Leaves Diarrhea, dysentery, malaria, and
snake bites [10, 16]
Leucas
aspera
(LA)
Lamiaceae Dronapushpi Whole
plant The decoction of whole plant of
Leucas aspera are used in malarial
[17, 18]
Demia
extensa
(DE)
Apocynaceae Dudheli Roots Antipyretic, expectorant, malarial
intermittent fever [16, 11]
Fagonia
Arabica
(FA) Zygophyllaceae Dhamaso Whole
plant Fever and malarial fever [19]
Table 2
Plant species with herbarium number and crude extract yields.
Plants Herbarium numbers Percentage Yield (%)
Methanol Hydro alcohol Aqueous
AE SU/DPS/Herb/58 18.8 24 31.2
BV SU/DPS/Herb/61 13 15 11.6
CC SU/DPS/Herb/65 18.8 24 24.2
AS SU/DPS/Herb/32 11.4 14.6 14.2
LA SU/DPS/Herb/18 8.6 12 12.2
PD SU/DPS/Herb/63/A 15.2 14.8 11.2
FA SU/DPS/Herb/63/B 9.2 8.6 8.2
Page 10/11
Table 3
Effect of methanol, hydro alcohol and aqueous extracts of selected medicinal plants on
Plasmodium
falciparum
lactate dehydrogenase (
Pf
LDH)
Plant species IC 50 (µg/mL) (MRC- 2) IC50 (µg/mL) (RKL − 9)
Methanol Hydro alcohol Water Methanol Hydro alcohol Water
AE 102.2** 274.2*** > 500* 274*** 313.8*** 315.7*
BV 24.65*** 39.42*** 172.1*** 28.71*** 62.22*** 110.2***
CC 103.3*** 142.6*** > 500*** 142.6*** 217.1*** > 500*
AS 78.11*** 88.87*** 348.5** 88.87*** 132.0*** 341.8**
LA - 431.1*** >500 431*** > 500* > 500
DE > 500 154.6** > 500* > 1000 478** > 500
FA > 1000 - > 500 - - -
Chloroquine 0.00382**
0.00221***
0.0264
0.00401***
Artemisinin
IC50 : 50% inhibitory concentration, the number of replicates was three, Signicance difference control
vs treatment *
p
≤ 0.05; **
p
≤ 0.01; ***
p
≤ 0.001.
Figures
Page 11/11
Figure 1
The effect of extracts of plants on schizont maturation of MRC-2 (A, B, C) and RKL- 9 (D, E, F) strains.
Results are expressed as mean ± SEM (n = 3). (A-Methanol extract, B- hydroalcohol extract, C- water
extract for MRC-2 strain) (D- Methanol extract, E- hydroalcohol extract, F- water extract for RKL-9 strain)
ResearchGate has not been able to resolve any citations for this publication.
Extracts of leaves and roots of Ocimum sanctum Linn. and leaves of Bauhinia variegata Linn. were used to evaluate their antimalarial effect against Plasmodium berghei in vivo in a typical 4 day test. Water and ether soluble extracts were administered orally to experimental mice along with placebo controls. On day 4 parasitaemia in control group of mice was 25.20% ± 9.44% while in mice treated with water soluble extracts of leaves and roots of Ocimum sanctum showed 2.80% ± 2.17% and 7.60% ± 5.32% infection respectively while in mice treated with water soluble extract of leaves of Bauhinia variegata showed 23.60% ± 13.35% infection.
In an effort to identify novel therapeutic alternatives for the treatment of malaria, the present study evaluated the antimalarial effect of the crude hydroalcoholic extract (HCE) from the leaves of Chenopodium ambrosioides L. For this purpose, the molecular affinity between the total proteins from erythrocytes infected with Plasmodium falciparum and HCE or chloroquine was evaluated by surface plasmon resonance (SPR). Subsequently, the plasmodicidal potential of HCE was assessed in a P. falciparum culture. Using BALB/c mice infected with Plasmodium berghei intraperitoneally (ip.), we evaluated the effects of ip. treatment, for three consecutive days (day 7, 8, and 9 after infection), with chloroquine (45 mg/kg) or HCE (5 mg/kg), considering the survival index and the parasitaemia. The groups were compared to an untreated control group that receives only PBS at the same periods. The results indicated that HCE could bind to the total proteins of infected erythrocytes and could inhibit the parasite growth in vitro (IC50 = 25.4 g/mL). The in vivo therapeutic treatment with HCE increased the survival and decreased the parasitaemia in the infected animals. Therefore, the HCE treatment exhibited a significant antiplasmodial effect and may be considered as a potential candidate for the development of new antimalarial drugs.
Malaria, caused by plasmodium parasite, is at the moment the highest cause of morbidity and mortality in the tropics. Recently, there is increasing efforts to develop more potent antimalarials from plant sources that will have little or no adverse effects. This study is aimed at investigating the in vivo mice antimalarial and in vitro antiplasmodial effects of two Meliaceae plants commonly used in Nigerian ethnomedicine as part of recipe for treating malaria infection: Chukrasia tabularis and Turraea vogelii. Hot water decoction and methanol extract of both plants were evaluated for their antimalarial activity in vivo using the mice model assay and in vitro using the parasite lactate dehydrogenase (pLDH) assay. The extracts were also assessed for toxicity with brine shrimp lethality assay and in mammalian cell lines using the neural red assay. The in vivo mice model antimalarial study showed that the methanol extract of the stem bark of C. tabularis exhibited the highest % chemosuppression (83.65 ± 0.66) at the highest dosage administered (800 mg/kg) when compared with chloroquine diphosphate, the standard reference drug which had a % suppression of 90.36 ± 0.04 (p < 0.05). The in vitro antiplasmodial study indicated that the aqueous extract of the stem bark of C. tabularis displayed good activity against Plasmodium falciparum chloroquine-sensitive (D6) strain (IC50 of 10.739 μg/mL) and chloroquine-resistant (W2) strain. Chloroquine and artemisinin had <0.163 and <0.0264, respectively.
-
![David J Newman]()
- Gordon M. Cragg
This contribution is a completely updated and expanded version of the four prior analogous reviews that were published in this journal in 1997, 2003, 2007, and 2012. In the case of all approved therapeutic agents, the time frame has been extended to cover the 34 years from January 1, 1981, to December 31, 2014, for all diseases worldwide, and from 1950 (earliest so far identified) to December 2014 for all approved antitumor drugs worldwide. As mentioned in the 2012 review, we have continued to utilize our secondary subdivision of a "natural product mimic", or "NM", to join the original primary divisions and the designation "natural product botanical", or "NB", to cover those botanical "defined mixtures" now recognized as drug entities by the U.S. FDA (and similar organizations). From the data presented in this review, the utilization of natural products and/or their novel structures, in order to discover and develop the final drug entity, is still alive and well. For example, in the area of cancer, over the time frame from around the 1940s to the end of 2014, of the 175 small molecules approved, 131, or 75%, are other than "S" (synthetic), with 85, or 49%, actually being either natural products or directly derived therefrom. In other areas, the influence of natural product structures is quite marked, with, as expected from prior information, the anti-infective area being dependent on natural products and their structures. We wish to draw the attention of readers to the rapidly evolving recognition that a significant number of natural product drugs/leads are actually produced by microbes and/or microbial interactions with the "host from whence it was isolated", and therefore it is considered that this area of natural product research should be expanded significantly.
Background In the face of chronic and emerging resistance of parasites to currently available drugs and constant need for new anti-malarials, natural plant products have been the bastion of anti-malarials for thousands of years. Moreover natural plant products and their derivatives have traditionally been a common source of drugs, and represent more than 30% of the current pharmaceutical market. The present study shows evaluation of anti-malarial effects of compound conessine isolated from plant Holarrhena antidysenterica frequently used against malaria in the Garhwal region of north-west Himalaya. Methods In vitro anti-plasmodial activity of compound was assessed using schizont maturation and parasite lactate dehydrogenase (pLDH) assay. Cytotoxic activities of the examined compound were determined on L-6 cells of rat skeletal muscle myoblast. The four-day test for anti-malarial activity against a chloroquine-sensitive Plasmodium berghei NK65 strain in BALB/c mice was used for monitoring in vivo activity of compound. In liver and kidney function test, the activity of alkaline phosphatase (ALP) was examined by p-NPP method, bilirubin by Jendrassik and Grof method. The urea percentage was determined by modified Berthelot method and creatinine by alkaline picrate method in serum of mice using ENZOPAK/CHEMPAK reagent kits. Results Compound conessine showed in vitro anti-plasmodial activity with its IC50 value 1.9 μg/ml and 1.3 μg/ml using schizont maturation and pLDH assay respectively. The compound showed cytotoxity IC50= 14 μg/ml against L6 cells of rat skeletal muscle myoblast. The isolated compound from plant H. antidysenterica significantly reduced parasitaemia (at 10 mg/kg exhibited 88.95% parasite inhibition) in P. berghei-infected mice. Due to slightly toxic nature (cytotoxicity = 14), biochemical analysis (liver and kidney function test) of the serum from mice after administration of conessine were also observed. Conclusion The present investigation demonstrates that the compound conessine exhibited substantial anti-malarial property. The isolated compound could be chemically modified to obtain a more potent chemical entity with improved characteristics against malaria.
Background Natural products could play an important role in the challenge to discover new anti-malarial drugs. In a previous study, Dicoma tomentosa (Asteraceae) was selected for its promising anti-plasmodial activity after a preliminary screening of several plants traditionally used in Burkina Faso to treat malaria. The aim of the present study was to further investigate the anti-plasmodial properties of this plant and to isolate the active anti-plasmodial compounds. Methods Eight crude extracts obtained from D. tomentosa whole plant were tested in vitro against two Plasmodium falciparum strains (3D7 and W2) using the p-LDH assay (colorimetric method). The Peters' four-days suppressive test model (Plasmodium berghei-infected mice) was used to evaluate the in vivo anti-plasmodial activity. An in vitro bioguided fractionation was undertaken on a dichloromethane extract, using preparative HPLC and TLC techniques. The identity of the pure compound was assessed using UV, MS and NMR spectroscopic analysis. In vitro cytotoxicity against WI38 human fibroblasts (WST-1 assay) and haemolytic activity were also evaluated for extracts and pure compounds in order to check selectivity. Results The best in vitro anti-plasmodial results were obtained with the dichloromethane, diethylether, ethylacetate and methanol extracts, which exhibited a high activity (IC50 ≤ 5 μg/ml). Hot water and hydroethanolic extracts also showed a good activity (IC50 ≤ 15 μg/ml), which confirmed the traditional use and the promising anti-malarial potential of the plant. The activity was also confirmed in vivo for all tested extracts. However, most of the active extracts also exhibited cytotoxic activity, but no extract was found to display any haemolytic activity. The bioguided fractionation process allowed to isolate and identify a sesquiterpene lactone (urospermal A-15-O-acetate) as the major anti-plasmodial compound of the plant (IC50 < 1 μg/ml against both 3D7 and W2 strains). This was also found to be the main cytotoxic compound (SI = 3.3). While this melampolide has already been described in the plant, this paper is the first report on the biological properties of this compound. Conclusions The present study highlighted the very promising anti-plasmodial activity of D. tomentosa and enabled to identify its main active compound, urospermal A-15-O-acetate. The high anti-plasmodial activity of this compound merits further study about its anti-plasmodial mechanism of action. The active extracts of D. tomentosa, as well as urospermal A 15-O-acetate, displayed only a moderate selectivity, and further studies are needed to assess the safety of the use of the plant by the local population.
Organic and aqueous extracts obtained from 14 Kenyan medicinal plants were screened for their antimalarial properties on two strains of Plasmodium falciparum (K1 chloroquine resistant and NF54 chloroquine sensitive). Dichloromethane extracts had the highest activities with IC50 ranging from 1.4 to 35.2 μg/ml. These extracts together with methanol extract of Turraea robusta were tested for their cytotoxicity properties in vitro on mammalian L6 cell line. The cytotoxicities ranged from > 90 to 0.34 μg/ml. Selectivity index (IC50 L6 cells/IC50P. falciparum) was also determined. Vernonia lasiopus had the highest selectivity index (SI) of greater than 10 while Warbugia ugandensis had the lowest SI of 0.24. This study suggests that V. lasiopus has a high potential for exploitation as a source of antimalarial agents.
- William Trager
- J B Jensen
Plasmodium falciparum can now be maintained in continuous culture in human erythrocytes incubated at 38°C in RPMI 1640 medium with human serum under an atmosphere with 7 percent carbon dioxide and low oxygen (1 or 5 percent). The original parasite material, derived from an infected Aotus trivirgatus monkey, was diluted more than 100 million times by the addition of human erythrocytes at 3- or 4-day intervals. The parasites continued to reproduce in their normal asexual cycle of approximately 48 hours but were no longer highly synchronous. They have remained infective to Aotus.
Caesalpinia crista L. (family Caesalpiniaceae) is a large scandent prickly evergreen shrub widely distributed in south-eastern Asia, Nigeria and Pacific region. The leaves are useful to cure skin infections, sore throat, intestinal worms, hepatic disorders, malaria and leprosy. Phytochemical investigation of a methanolic extract of the leaves of C. crista afforded two new cheilanthane-type tricarbocyclic sesterterpenoids characterized as 4α,4β,8β,13β-tetramethyl-14-(17-methyl-pent-14-enyl)-perhydrophenanthrene-11α-ol-22-oic acid (1, cristasesterterpenoic acid) and 4α,4β,8β,10β,13β-pentamethyl-14-(-17-methylpent-14-enyl)-perhydrophenanthrene-3β-olyl-O-β-D-glucopyranoside (2, cristasesterterpinol glucoside). The structures of these compounds were elucidated on the basis of spectral data analysis and chemical reactions.
Source: https://www.researchgate.net/publication/344717821_Antiplasmodial_Potential_of_Traditional_Phytotherapy_of_Some_Remedies_Used_in_The_Treatment_of_Malaria_in_India
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