International Journal of Pharmaceutical Chemistry and Analysis 2023;10(3):209–214
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International Journal of Pharmaceutical Chemistry and
Analysis
Journal homepage: https://www.ijpca.org/
Original Research Article
In vitro antileishmanial evaluation of Vernonia Brachycalyx leaf latex extract
against two leishmania species
Alemu Tadesse Feroche
1, *
1 Dept. of Pharmaceutical Chemistry and Pharmacognosy, Addis Ababa University, School of Pharmacy, Addis Ababa, Ethiopia
ARTICLE INFO
ABSTRACT
Article history:
Received 08-08-2023
Accepted 05-09-2023
Available online 15-09-2023
Background: Leishmaniasis is a major public health problem, and the alarming spread of parasite
resistance has increased the importance of discovering new therapeutic products. In the present study,
the antileishmanial activity of the methanolic extract of the leaf latex obtained from the Ethiopian plant
Vernonia brachycalyx O. H. (family Asteraceae) was evaluated by in vitro testing against Leishmania
aethiopica and L. donovani.
Materials and Methods: Antileishmanial activity test was carried out using the Alamar Blue assay on
promastigotes and axenic cultured amastigotes of L. aethiopica and L. donovani clinical isolates, and cell
viability was fluorometrically determined. Amphotericin B was used as a positive control, and 1% dimethyl
sulfoxide (DMSO) and the media were employed as a negative control.
Moreover, preliminary phytochemical analysis of the extracts was performed.
Results: Results of the study indicated that the latex possesses good activity against both parasites, with
IC50 values of 6.82 ± 0.18 and 6.34 ± 0.20µg/ml against promastigotes and 3.53 ± 0.33 and 2.61 ±
0.907µg/ml against axenically cultured amastigotes of L. aethiopica and L. donovani, respectively. The
latex demonstrated selectivity indices (SIs) of 15.27 and 16.42 against promastigotes and 29.50 and
39.90 against axenically cultivated amastigotes of L. aethiopica and L. donovani. While, amphotericin
B demonstrated SIs of 7.91 and 8.23 against promastigotes and 7.45 and 7.73 against axenically cultured
amastigotes of and L. donovani, respectively. Phytochemical screening demonstrated that the latex contains
flavonoids, tannins, cardiac glycosides, terpenoids, saponins, alkaloids, and steroids.
Conclusion: The findings of this investigation attest that the latex of V. brachycalyx possesses promising
antileishmanial activity against L. aethiopica and L. donovani, warranting further investigations into the
active constituents.
Keywords:
Antileishmanial activity
in vitro
leishmania aethiopica
leishmania donovani
Vernonia brachycalyx
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1. Introduction
considered an emerging disease with high morbidity and
mortality in the tropics and subtropics (Ornellas-Garcia et
al., 2023). Leishmania is characterized by both its diversity
and its complexity. Depending on the parasite strain(s)
involved in the pathogenesis and host immune response,
it can cause clinical symptoms ranging from mild, selflimiting skin lesions to visceral disease. (Loría-cervera et
al., 2014; Tariku, 2008). There are several different forms of
leishmaniasis in humans. The three main clinical forms are
cutaneous, mucocutaneous, and visceral leishmaniasis. A
Leishmania is a disease caused by a single-celled parasite
of the genus Leishmania, which is spread by the bite
of several species of fireflies (subfamily Phlebotomine).
Although not bearing the same country name as malaria,
Leishmaniasis continues to have a major impact on a
significant portion of the world’s population and is currently
* Corresponding author.
E-mail address: Alemu.tadesse@aau.edu.et (A. T. Feroche).
https://doi.org/10.18231/j.ijpca.2023.035
2394-2789/© 2023 Innovative Publication, All rights reserved.
209
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Feroche / International Journal of Pharmaceutical Chemistry and Analysis 2023;10(3):209–214
fourth, less common form is diffuse cutaneous leishmaniasis
(Pagheh et al., 2023).According to the World Health
Organization, the incidence of leishmaniasis has increased
worldwide in recent decades. Leishmania is endemic in
200 countries in Asia, Africa, the Americas, and the
Mediterranean region (WHO, 2021). Leishmaniasis causes
2 million disability-adjusted life years in 90 countries,
mostly in the developing world (Desjeux, 2004). In
Ethiopia, cutaneous leishmaniasis is mainly caused by the
endemic species L. aethiopica and rarely by L. tropica and
L. major. It is found almost everywhere in the country at
altitudes between 1500 and 2700 meters (Hailu et al., 2007).
Visceral leishmaniasis (VL) is distributed in lowlands and
semi-deserts (below 1500 m altitude) with varying degrees
of prevalence. To date, cases of VL have been reported in
at least 40 localities, with an estimated annual burden of
2000–4500 cases in Ethiopia (Ministry of Health, 2013).
Treatments for human leishmaniasis include pentavalent
antimonial, amphotericin B, pentamidine, miltefosine, and
paromomycin. Due to frequent side effects and increasing
drug resistance, these drugs are not effective. Therefore,
the need for better, safer, and more effective drugs is
of utmost importance (Feroche et al., 2022). This poses
a need for new antileishmanial drugs to stop the spread
of the disease.In addition, to find new lead compounds
and/or disease-fighting drugs to investigate, researchers
are focusing on natural compounds used to treat parasitic
infections, including leishmaniasis (Ayalew et al. 2018).
The use of plants as alternative medicine in Ethiopia
has been an ancient practice for centuries. In fact, in
Ethiopia, for 70–80% of the population and about 90% of
livestock, traditional medicines are used as the main source
of treatment (Chekole et al., 2017). Therefore, in this study,
the in vitro antileishmanial activity, phytochemical analysis,
and in vitro cytotoxicity studies of the 80% methanolic
Vernonia brachycalyx latex extract were investigated. 1–3
2. Materials and Methods
2.1. Plant material collection
Fresh latex of V. brachycalyx was collected from the Bale
Mountains, Oromia Regional State, Ethiopia, in June 2021.
The identification and authenticity of the plant material
were confirmed by Mr. Melkamu Wondafrash, National
Herbarium, Department of Plant Biology and Biodiversity
Management, College of Natural and Computational
Sciences, Addis Ababa University. The resulting latex is
left to dry in the shade. The dried latex is then ground
into a coarse powder and stored in a desiccator at room
temperature until further use.
2.2. Preparation of latex extract
The latex is collected from the leaves of V. brachycalyx by
cutting the leaves at the bottom and letting the latex drip
onto a plastic sheet. The latex is then allowed to air dry
for 3 days to form a light powder. 4–11 The dry material was
then weighed and stored in a refrigerator at 4◦ C until use.
Powdered dry latex (30 g) was soaked with 80% methanol
(3 × 0.5 L, 72 h each). The extract was then filtered, and the
methanol in the filtrate was removed by a rotary evaporator
to obtain 19 g of brown powder.
2.3. Cell culture promastigote culture
The L. aethiopica and L. donovani were grown in
tissue culture flasks containing RPMI 1640 medium
(Gibco, Invitrogen Co., UK) supplemented with 10% heatinactivated fetal bovine serum (HIFCS) (Gibco, Invitrogen
Co., United Kingdom) and 100 IU penicillin/ml -100µg/ml
streptomycin solution (Sigma Chemical Co., St. Louis,
USA) at 220 C for L. aethiopica and 260 C L. donovani
(Nigatu et al., 2021). Cell-free medium was used to culture
the parasites in vitro and set up a test system to determine
the IC50 value of the extract.
2.4. Axenic cultured amastigote
Axenically cultured amastigotes were cultured according
to the methods described for L. aethiopica and L.
donovani (Nigatu et al., 2021) with minor modifications.
Late stationary phase promastigotes were centrifuged
and resuspended in Hank’s balanced salt 199 medium,
supplemented with 20% heat-inactivated fetal bovine serum
(HIFCS), 2 mM L-glutamine, penicillin 50 IU/mL and
streptomycin 50µg/mL; pH was adjusted to 5.5 (for both
strains) with 0.10N HCl. The cells were then incubated at
31◦ C (for L. aethiopica) or 37◦ C (for L. donovani) in a
humidified 5% CO2 incubator for 7 days.
2.5. In vitro antileishmanial test
All experiments were performed in triplicate. Briefly, serial
dilutions of Vernonia brachycalyx were prepared. Final
concentrations of 100, 50, 25, 12.5, and 6.25 µg/mL
were performed to establish adequate dose titration and to
determine the IC50 value. Plant latex was added to a 96well microtiter plate containing 100 µL of complete culture
medium to achieve a final concentration of 100 µg/mL
of each. Then, a 100µL suspension of the parasite (3.5
× 106 promastigotes of L. aethiopica and L. donovani)
obtained from previous cultures were added to each well.
The parasites were then incubated for 72 h at room
temperature for the promastigotes of the two strains, at
31◦ C and 37◦ C for axenic amastigote cultures of L.
aethiopica and L. donovani, respectively, in the presence
of different concentrations of the extract. Then, resazurin
(0.125 mg/mL) was added to the 20 µL suspension (10%
of the total volume of each well). The mixture is covered
with aluminum foil and left at the above temperature.
Fluorescence intensity was measured using a Victor 3
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Feroche / International Journal of Pharmaceutical Chemistry and Analysis 2023;10(3):209–214
multilabel counter (PerkinElmer, MA, USA) at an excitation
wavelength of 544 nm and an emission wavelength of 590
nm. The trial was performed in triplicate and compared with
negative controls (1% DMSO and medium alone) and a
reference drug (amphotericin B, Sigma-Aldrich, Germany).
During the assay, cell viability was monitored by measuring
the fluorescence signal. The fluorescence intensity produced
is proportional to the number of viable cells (Tewabe et al.,
2019).
2.6. Study of cytotoxicity in THP-1 monocytes
THP-1 cells were cultured in RPMI 1640 medium (SigmaAldrich, Co., St. Louis, USA) supplemented with 10%
HINBCS, 100 IU/ml penicillin, and 100 µg/ml streptomycin
at 37◦ C in a 5% CO2 humidified incubator (Thermo
Scientific, USA). In 96-well plates, THP-1 monocytes were
plated at a density of 4 × 104 cells per well (in a volume
of 200 µl) with or without latex extract, and the plates were
incubated at 37 ◦ C with 5% CO2 for 72 h. Then Alamar
Blue was added. 12–22 During the final 3 h of incubation,
cell viability was measured by fluorescence measurement
as described previously (A. T. Feroche et al., 2021).
selective agents for the treatment of other tropical diseases
caused by protozoa (Ribeiro TG. et al., 2014).
Table 1: Antipromastigote activity of V. brachycalyx against
Leishmania aethiopica and Leishmania donovani.
Test substance
Latex extract
AmB (reference)
DMSO (NC)
Media alone (NC)
Notes: Values are expressed as mean ± SD; n = 3. x Effective concentration
required to achieve 50% growth inhibition in µg/mL; y no effect.
Abbreviations: NC, negative control; DMSO, dimethyl sulphoxide. AmB,
Amphotericin B
Table 2: Effects of V. brachycalyx on axenically cultured
amastigotes of Leishmania aethiopica and Leishmania donovani
and THP-1 monocytes.
Antipromastigote
activityIC50 (µg/mL) x
Test
substance
2.7. Phytochemical screening
V. brachycalyx leaf latex extract in methanol has
been used to evaluate the presence or absence of
plant components such as alkaloids, saponins, tannins,
anthraquinones, flavonoids, steroids, cardiac glycosides,
resins, phenolic compounds, phlobatannin, comarin, and
terpenoid. Standard methods for preliminary phytochemical
analysis were used (Alamzeb et al., 2013; Thusa and Mulmi,
2017). 23–29
Antipromastigote activityIC50
(µg/mL) x
L. aethiopica
L. donovani
6.82 ± 0.18
6.34 ± 0.20
1.29 ± 0.08
1.24 ± 0.01
0.00y
0.00y
0.00y
0.00y
Latex
extract
AmB
(reference)
DMSO
(NC)
Media alone
(NC)
Cytotoxic
Effect in
THP-1
LC50
(µg/mL)
L. aethiopica
3.53 ± 0.33
L. donovani
2.61 ± 0.907
1.37 ± 0.05
1.32 ± 0.15
0.00y
0.00y
104.13 ±
0.94
10.21 ±
0.25
00
0.00y
0.00y
00
Notes: Values are expressed as mean ± SD; n = 3. x Effective concentration
required to achieve 50% growth inhibition in µg/mL; y no effect.
2.8. Data analysis
Antileishmanial activity (IC50 ) values were calculated from
sigmoid inhibitor dose-response curves using the computer
software GraphPad Prism 8.0.1.244 (GraphPad Software,
Inc., CA, USA) and Microsoft Excel. Values are expressed
as the mean standard deviation of triplicate experiments.
3. Result and Discussion
3.1. Antileishmanial activity
Current alternative treatment options for leishmaniasis are
considered inadequate because these drugs are associated
with high toxicity and high cost, and parasite resistance
to these drugs is increasing (Tasdemir et al., 2006).
In this context, strategies to identify a new compound
with lower toxicity and better efficacy may be useful, as
they are considered highly desirable. As an alternative to
chemotherapy in the treatment of parasitic diseases, herbal
medicines have received much attention. Furthermore,
natural products have potential advantages as novel and
Table 3: Phytochemical analysis of an 80% methanol latex extract
of V. brachycalyx
Phytochemicals
Alkaloid
Cardiac glycosides
Flavonoids
Saponins
Steroids
Tannins
Terpenoids
Cardiac glycosides
V. brachycalyx latex extract
+++
+++
+++
+++
+++
+++
+++
+++
Therefore, this study was conducted with the aim of
finding compounds that inhibit the in vitro growth of two
Leishmania parasites, L. aethiopica and L. donovani. First,
the leaf latex of V. brachycalyx was tested for its growth
inhibition against L. aethiopica and L. donovani, as several
plants of the genus Vernonia have been reported to have
antiprotozoal activity (C.N. Muthaura et al., 2007). Indeed,
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plant latex showed strong inhibitory activity on the flagellate
and extracellular premature stages of L. aethiopica and L.
donovani, with IC50 values of 6.82 ± 0.18 and 6.34 ± 0.20
respectively (Table 1). However, these results are considered
preliminary, as Leishmania promastigote is more sensitive
to drug-induced effects than amastigotes (Y. Baquedano
et al., 2016). Furthermore, the clinical manifestations
of the disease in humans are related to intracellular
amastigotes, which are the evolved form of vertebrate
parasites, and not to extracellular promastigotes. Therefore,
the latex was further tested against axenic amastigotes of
both pathogenic parasites.Tests showed that the latex had
better activity than that observed for the promastigotes
of L. aethiopica and L. donovani, with IC50 values of
3.53 ± 0.33 and 2.61 ± 0.907µg/mL respectively (Table
2).At higher concentrations, leaf latex showed the highest
inhibitory activity against promastigote and amastigote of
L. aethiopica and L. donovani. This indicates that there is
a direct correlation between the concentration of the latex
extract and the percentage inhibition.The antileishmanial
effect of the reference drug, amphotericin B, was also
determined for comparison purposes in clinical isolates and
strains of L. donovani and L. aethiopica.For comparison, the
antileishmanial effect of the reference drug, amphotericin B,
was determined both in clinical isolates of L. aethiopica and
L. donovani.Even so, the leaf latex extract was less active
than amphotericin B, having IC50 values of 1.29 ± 0.08
µg/mL and 1.24 ± 0.01 µg/mL on promastigotes (Table 1)
and IC50 values of 1.37 ± 0.05µg/mL and 1.32 ± 0.15
µg/mL on axenically cultured amastigote of L. aethiopica
and L. Donovani, respectively (Table 2).
4. Phytochemical Analysis
The results of various phytochemical screening tests
obtained during the experiment were flavonoids, tannins,
cardiac glycosides, terpenoids, saponins, quinones,
alkaloids, and steroids (Table 3). Numerous studies
performed on several plant secondary metabolites have
reported their potential growth inhibitory effects on
Leishmania spp. These include alkaloids (Henriques et
al., 2001), steroids (Sartorelli et al., 2007), polyphenols
(Bodiwala et al., 2007), tannins (Kolodziej et al., 2001),
flavonoids (Tasdemir et al., 2006), anthraquinones (ChanBacab et al., 2001), saponins (Maes et al., 2004), and
terpenoids (Kayser et al., 2003). The biological activities
observed in this V. brachycalyx may be due to individual
groups of compounds present in the plant or to a synergistic
effect caused by individual compounds.
4.1. In vitro cytotoxicity studies
A large proportion of the world’s population does not have
access to standard medicines and is dependent on traditional
natural medicines. To complement this growing interest in
alternative treatment plans and ensure safe therapeutic use
of herbal medicines, their toxicity needs to be investigated.
Toxicity testing indicates how safe the herbal medicine
is, and the results of these tests are very important for
further in vivo studies. In this study, cytotoxicity tests
were performed in vitro to evaluate the safety of the latex
extract.The selectivity index (SI) is an important tool for
describing the safety of biologically active compounds. The
SI determined for the tested substances is expressed as the
ratio between cytotoxicity (LC50 value on THP-1 cells)
and activity (IC50 value on L. aethiopica or L. donovani
amastigote). The latex exhibited a selectivity index (SI)
of 15.27 and 16.42 for promastigote and 29.50 and 39.90
for axenically cultured amastigote of L. aethiopica and L.
donovani. While amphotericin B exhibited SIs of 7.91 and
8.23 for promastigotes and 7.45 and 7.73 for axenically
cultured amastigotes of L. aethiopica and L. donovani,
respectively. The latex extract showed a much higher SI
than the reference drug, amphotericin B, indicating its high
selectivity for Leishmania parasites.
In the present study, the latex extract was twice as
selective as amphotericin B on the promastigotes of L.
aethiopica or L. donovani and four times more selective
than amphotericin B on the amastigotes of L. aethiopica or
L. donovani. The results obtained in this study are much
higher than the results of other similar studies conducted
with the same genus of Vernonia on Leishmania aethiopica
(Azeb T. et al., 1993). Furthermore, it was higher than
that reported for latex extracts of A. rugosifolia (IC50
= 31.21 ± 0.01 µg/ml and IC50 = 24.5 ± 0.24 µg/ml)
(Chemeda et al., 2022) on the promastigote and amastigote
stages of the parasite.Although the potency of the tested
substances against intracellular amastigotes was lower than
that of the reference drug, their low toxicity to THP-1
cells and their high SI represent a great advantage over the
reference drug. Compounds with SI values above 20 are
ideal candidates for further development of antileishmanial
agents (Nwaka S. and Hudson A., 2006).For axenically
cultured amastigotes in the present work, the latex extract
exhibited a significantly greater SI value than 20. According
to this criterion, the latex extract of V. brachycalyx may
serve as a model for developing safer and more effective
antileishmanial agents. In addition, latex extracts have the
added advantage of being used therapeutically in their own
right as they are active components of many common herbal
medicines that have long been used as antimalarial agents
(C.N. Muthaura et al., 2007).
5. Conclusion
The results of this study demonstrated that the methanol
extract of the leaf latex of V. brachycalyx showed
promising activity, with a direct correlation between the
latex extract concentration and the percentage inhibition
against promastigotes and axenically cultured amastigotes
Feroche / International Journal of Pharmaceutical Chemistry and Analysis 2023;10(3):209–214
of L. aethiopica and L. donovani, respectively. This could
be explained by the existence of secondary metabolites such
as flavonoids and terpenoids, which have previously been
reported to have true antileishmanial activity. Furthermore,
the extract showed much higher SI values than the reference
drug, amphotericin B, which showed high selectivity
for Leishmania parasites. However, the leaf latex extract
was less active than amphotericin B on promastigotes
and amastigotes of L. aethiopica and L. donovani,
respectively.The promising activity profile of the methanol
extract of V. brachycalyx leaf latex, together with its
relative safety margin, requires further study of the plant’s
phytochemicals to isolate the compounds responsible for its
antileishmanial activity. Plant bioactive compounds can be
isolated and used as effective anti-leishmanial agents or as
starting points for the development of safer, more effective,
and more economical leishmaniasis treatment options.Latex
extracts have also been tested for phytochemicals. The
plant components found were flavonoids, tannins, cardiac
glycosides, terpenoids, saponins, quinones, alkaloids, and
steroids.
6. Data Sharing Statement
The corresponding author (Alemu Tadesse) will make the
data used to support the findings of this study available upon
reasonable request from the school of pharmacy at Addis
Ababa University. Email: donganegn@gmail.com
7. Author Contributions
Mr. Alemu Tadesse contributed to the research’s design,
synthesis, evaluation, and laboratory work, as well as the
analysis of results and manuscript writing.
8. Abbreviations
CL, Cutaneous Leishmaniasis; DMSO, Dimethyl
Sulfoxide; IC50 , the half maximal inhibitory concentration;
MCL, Mucocutaneous Leishmaniasis; VL, Visceral
Leishmaniasis; DCL, Diffuse Cutaneous Leishmaniasis;
PKDL, Post kala-azar Dermal Leishmaniasis; USA, United
States of America; HIFCS, Heat Inactivated Fetal Calf
Serum.
9. Acknowledgments
My heartfelt thanks to Addis Ababa University,
the Department of Microbiology, Immunology, and
Parasitology, and the Department of Microbial Cellular and
Molecular Biology for providing instruments, chemicals,
and laboratory strains for completing this study.
10. Source of Funding
No funding available.
213
11. Declaration of Interest
The author declares that there is no conflict of interest.
References
1. Chekole G. Ethnobotanical study of medicinal plants used against
human ailments in Gubalafto District, Northern Ethiopia. J Ethnobiol
Ethnomed. 2007;13(1):55.
2. Chemeda G, Bisrat D, Yeshak MY, Asres K. In vitro antileishmanial
and anti-trypanosomal activities of plicataloside isolated from the
leaf latex of Aloe rugosifolia Gilbert & Sebsebe (Asphodelaceae).
Molecules. 2022;27:1400.
3. Feroche AT, Awoke D, Hailu A.
Anti-Leishmanial Activity
Investigation of Some New Quinazolinone Derivatives. Uttar Pradesh
J Zool. 2022;4(5):56–71.
4. Hailu A, Gebre-Michael T, Berhe N, Meshesha B. The epidemiology
and ecology of health and disease in Ethiopia. Addis Ababa, Ethiopia:
Sa Books; 2006. p. 615–49.
5. Henriques AT, Bou-Habib, Saraiva EM. Antileishmanial activity
of an indole alkaloid from Peschiera australis. Antimicrob Agennt
Chemother. 2001;45:1349–54.
6. Kayser O, Kiderlen AF, Croft S. Natural Products as potential
antiparasitic drugs . Stud Nat Prod Chem. 2007;26:779–848.
7. Kolodziej H, Kayser O, Kiderlen AF, Ito H, Hatano T, Yoshida T, et al.
Antileishmanial activity of hydrolyzable tannins and their modulatory
effects on nitric oxide and tumour necrosis factor-alpha release in
macrophages in vitro. Planta Med. 2001;67:825–57.
8. Loría-Cervera EN, Andrade-Narváez F. Animal models for the
study of leishmaniasis immunology. Rev Inst Med Trop Sao Paulo.
2014;56(1):1–11.
9. Maes L, Berghe DV, Germonprez N, Quirijnen L, Cos P, De Kimpe
N. In vitro and in vivo activities of a triterpenoid saponin extract
(PX- 6518) from the plant Maesa balansae against visceral leishmania
species. Antimicrob Agent Chemother. 2004;48:130–66.
10. Ministry of Health, Ethiopia. Guideline for diagnosis, treatment and
prevention of Leishmaniasis in Ethiopia. 2nd edition June, Addis
Ababa; 2013. Available from: http://repository.iifphc.org/handle/
123456789/445.
11. Garcia UO, Cuervo P, Fl RG. Malaria and leishmaniasis: Updates on
co-infection. Front Immunol. 2023;14:1122411.
12. Desjeux P. Leishmaniasis: current situation and new perspectives.
Comp Immun Microbiol Infect Dis. 2004;27(5):305–18.
13. Pagheh AS, Fakhar M, Mesgarian F, Rahimi-Esboei B, Badiee F.
Incidence trend of rural cutaneous leishmaniasis. J Mazandaran Univ
Med Sci. 2013;23(104):27–33.
14. Ribeiro TG. Antileishmanial activity and cytotoxicity of Brazilian
plants. Experimental Parasitology. 2014;143:60–68.
15. Nwaka S, Hudson A. Innovative lead discovery strategies for tropical
diseases. Nat Rev Drug Dis. 2006;5(11):941–55.
16. Sartorelli P, Andrade SP, Melhem M, Prado FO, Tempone AG.
Isolation of antileishmanial sterol from the fruits of Cassia fistula using
bioguided fractionation. Phytother Res. 2007;21:644–91.
17. Tariku Y. in vitro efficacy study of some selected medicinal plants
against leishmania spp. M.Sc. thesis School of Graduate Studies.
Ethiopia; 2008.
18. Tasdemir D, Kaiser M, Brun R, Schmidt YV, Tosun TJ, Rüedi F,
et al. Antitrypanosomal and leishmanicidal activities of flavonoids and
their analogues: in vitro, in vivo, structure-activity relationship, and
quantitative structure-activity relationship studies. Antimicrob Agents
Chemother. 2006;50(4):1352–64.
19. Tewabe Y, Kefarge B, Belay H, Bisrat D, Hailu A, Asres K, et al.
Antipromastigotes Evaluation of the Leaf Latex of Aloe macrocarpa,
Aloin A/B, and Its Semisynthetic Derivatives against Two Leishmania
Species. Evid Based Comp Alternat Med. 2019;p. 4736181.
20. Thusa R, Mulmi S. Analysis of phytoconstituents and biological
activities of different parts of Mahonia nepalensis and Berberis
aristata. Nepal J Biotechnol. 2017;5:5–13.
214
Feroche / International Journal of Pharmaceutical Chemistry and Analysis 2023;10(3):209–214
21. Global Leishmaniasis Surveillance. A Baseline for the 2030 Road
Map; 2021. Available from: https://www.who.int/publications/i/item/
who-wer9635-401-419.
22. Baquedano Y, Alcolea V, Toro MA, et al.
Novel heteroaryl
selenocyanates and diselenides as potent antileishmanial agents.
Antimicrob Agents Chemother. 2016;60(6):3802–12.
23. Alamzeb M, Khan MR, Ali S, Shah SQ, Mamoon UR. Antimicrobial
properties of extracts and compounds isolated from Berberis
jaeschkeana. Bangladesh J Pharmcol. 2013;8(2):107–9.
24. Feroche AT, Derese K. In Vitro Antiprotozoal Activity of the Leaf
Latex of Caralluma speciosa Against Two Leishmania Species. Ethiop
Pharm J. 2021;37:153–60.
25. Ayalew H, Tadesse S, Lindemann DB, Tewabe P, Hailu Y. Vitro
Antipromastigotes Activity of Essential Oil of the Leaves of
Discopodium pennnervium Hochst. J Exp Pharmacol. 2018;34:75–
80.
26. Tadesse A, Gebrehiwot A, Ares K. In vitro activity of Vernonia
amygdalina on Leishmania aethopica. Ethiop Med J. 1993;31(3):183–
9.
27. Bodiwala HS, Singh G, Singh R, Dey CS, Sharma SS, Bhutani KK,
et al. Antileishmanial amides and lignans from Piper cubeba and Piper
retrofractum. J Nat Med. 2007;61:418–439.
28. Muthaura CN, Rukunga GM, Chhabra SC, Mungai GM, Njagi
ENM. Traditional phytotherapy of some remedies used in treatment
of malaria in Meru district of Kenya. South African J Botany.
2007;73:402–11.
29. Baquedano Y, Alcolea V, Toro MA. Novel Heteroaryl Selenocyanates
and Diselenides as Potent Antileishmanial Agents. Antimicrob Agents
Chemother. 2001;60(6):3802–12.
Author biography
Alemu Tadesse Feroche, –
https://orcid.org/0000-0002-8086-6925
Cite this article: Feroche AT. In vitro antileishmanial evaluation of
Vernonia Brachycalyx leaf latex extract against two leishmania species.
Int J Pharm Chem Anal 2023;10(3):209-214.