Prophylactic and curative activities of extracts from Warburgia ugandensis Sprague (Canellaceae) and Zanthoxylum usambarense (Engl.) Kokwaro (Rutaceae) against Plasmodium knowlesi and Plasmodium berghei

doi:10.1016/j.jep.2010.04.034

Journal of Ethnopharmacology

Volume 130, Issue 1, 6 July 2010, Pages 158-162

https://doi.org/10.1016/j.jep.2010.04.034 Get rights and content

Abstract

Ethnopharmacological relevance

This work reports the anti-plasmodial activities of Warburgia ugandensis and Zanthoxylum usambarense commonly used as phytomedicines against malaria by some Kenyan communities.

Aim of study

To determine the anti-plasmodial activities of extracts from Warburgia ugandensis and Zanthoxylum usambarense against Plasmodium knowlesi and Plasmodium berghei.

Materials and methods

Eight plant extracts were screened for in vitro anti-plasmodial activity against Plasmodium knowlesi, in a 96-well plate incubated at 37 °C on a RPMI culture medium supplemented with baboon serum. Of the eight, three were investigated for prophylactic and curative activities in BALB/c mice against drug-sensitive Plasmodium berghei in a 4-day test at a dose rate of 200 mg/kg/day.

Results

Inhibitory concentrations (IC₅₀) values of between 3.14 and 75 μg/ml, up to 69% chemosuppression of parasites growth and over 80% survivorship of treated mice were observed.

Conclusion

The two medicinal plants, Warburgia ugandensis and Zanthoxylum usambarense possess bioactive compounds against malaria parasites and could be exploited for further development into malaria therapy.

Graphical abstract

Introduction

Malaria is the most widespread haemoparasitic disease in the world, with an estimated incidence of about 500 million clinical cases annually and a corresponding annual mortality of up to 2.7 million (WHO, 1998). The high cost of effective treatment is one of the most important factors hampering malaria control efforts. Herbal medicine is used by local communities in the developing world to treat various infectious diseases (Bidla et al., 2004). In Africa, about 75% of the population either does not have access to or cannot afford conventional medicine and therefore resort to traditional medicine for treating malaria (Azas et al., 2001, Bourdy et al., 2004). Warburgia ugandensis Sprague (Canellaceae) and Zanthoxylum usambarense (Engl.) Kokwaro (Rutaceae) are commonly used as traditional medicine by many communities in Kenya (Njoroge and Bussmann, 2006). Both plants grow in highland areas especially in dry forests around Nairobi, Masaai Mara, Samburu and Southwest of Mt. Kenya (Nanyingi et al., 2008). Hot and cold decoctions from leaves and stem barks of Warburgia ugandensis are culturally used to treat tooth decay, asthma and bronchitis while Zanthoxylum usambarense is used to treat malaria, upper respiratory tract infections, tooth decay and sore gums (Nanyingi et al., 2008, Njoroge and Bussmann, 2006). Phytochemical studies on Zanthoxylum usambarense revealed that it contains alkaloids of tetrahydroprotoberberine type (Kato et al., 2007). Moreover, its in vitro antimalarial activity against Plasmodium falciparum has been reported (Kirira et al., 2006). This study describes the activity of extracts from Warburgia ugandensis and Zanthoxylum usambarense against the fifth human malaria parasite (White, 2008), Plasmodium knowlesi and the rodent malaria parasite Plasmodium berghei. Plasmodium knowlesi is widely distributed in Malaysia, the Philippines, Myanmar and Thailand, and can also be fatal, presenting an urgent need for more focused investigations on its control (Tek et al., 2008, Luchavez et al., 2008).

Section snippets

Plants

Fresh stem barks from Warburgia ugandensis and Zanthoxylum usambarense were collected from Oloolua Forest, Ngong Division of Kajiado District, Kenya and authenticated by a plant taxonomist, at the National Museums of Kenya (NMK). The plant parts collected were those used in traditional preparations as guided by a local herbalist. Voucher specimen number K4/H27/07 was deposited at the East African herbarium of the National Museums of Kenya.

Plant extracts

Forty grams (40 g) of air-dried and pulverized stem barks

In vitro activities of Warburgia ugandensis and Zanthoxylum usambarense against Plasmodium knowlesi

Anti-plasmodial activities of stem bark extracts from Warburgia ugandensis and Zanthoxylum usambarense were performed against Plasmodium knowlesi blood stage parasites. Warburgia ugandensis extracts of non-polar solvents were more inhibitory to Plasmodium knowlesi than its aqueous extract (Table 1), with the chloroform extract being the most active (IC₅₀ = 3.14 μg/ml). In contrast, the aqueous extract from Zanthoxylum usambarense was remarkably active (IC₅₀ = 6.04 μg/ml), with all organic solvents

In vitro anti-plasmodial activities and IC₅₀ values

The activity of the aqueous extract from Zanthoxylum usambarense on Plasmodium knowlesi (IC₅₀ = 6.04 μg/ml) observed in this study compares well with the findings of another study in which the IC₅₀ value for the aqueous extract from the same plant fell between 6 and 15 μg/ml against both chloroquine-sensitive and resistant Plasmodium falciparum strains, while that of methanolic extract was found to be lower than 6 μg/ml (Kirira et al., 2006). These findings are consistent with the results of our

Conflict of interest

The authors declare that there is no conflict of interest.

Acknowledgements

This investigation received financial support under the Multilateral Initiative on Malaria (MIM) Project ID A50075 through the UNICEF/UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases (TDR).

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Ethnobotanical uses, phytochemistry and pharmacology of pantropical genus Zanthoxylum L. (Rutaceae): An update
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The aqueous extract from Z. usambarense (Engl.) Kokwaro was remarkably active against Plasmodium knowlesi and Plasmodium berghei in both in vitro and in vivo IC50 = 6.04 g/mL (Were et al., 2010). These results suggest that the antiplasmodial activity of Z. usambarense is due mainly to polar substances.
Plants have been used in various parts of the world to treat various diseases. The genus Zanthoxylum L. (Rutaceae) is the second largest genus of this family and comprises approximately 225–549 species distributed in the tropical and temperate regions of the world. Plants of this genus are trees and shrubs with various applications in folklore medicine for food, medicine, construction, and other uses.
The goal of this review is to give an updated data on the ethnobotanical applications, phytochemistry, and pharmacology of the Zanthoxylum species to investigate their medicinal potential and identify research gaps for future research studies.
Data was obtained through a systematic search of published literature and online databases such as Google Scholar, Web of Science, PubMed, Science Direct, and Sci-Finder. The botanical names were confirmed using the World Flora Online and chemical structures were drawn using the ChemBio Draw Ultra Version 14.0 Software.
The Zanthoxylum species have a wide use in different parts of the continents as a remedy for various diseases such as digestive diseases, gastrointestinal disorders, venereal diseases, respiratory diseases, rheumatism, bacterial diseases, viral, and other diseases. Various parts of the plant comprising fruits, seeds, twigs, leaves, oils, and stems are administered singly or in the form of decoction, infusion, powder, paste, poultice, juice, or mixed with other medicinal plants to cure the disease. More than 400 secondary metabolites have been isolated and characterized in this genus with various biological activities, which comprise alkaloids, flavonoids, coumarins, lignans, alcohols, fatty acids, amides, sesquiterpenes, monoterpenes, and hydrocarbons. The crude extracts, fractions, and chemical compounds isolated from the genus have demonstrated a wide range of biological activities both in vivo and in vitro, including; anti-cancer, antimicrobial, anti-sickling, hepatoprotective, antipyretic, antitumor, and other pharmacological activities.
This genus has demonstrated an array of phytoconstituents with therapeutic potential. The ethnobotanical uses of this genus have been confirmed in modern pharmacological research. This genus is a potential source for modern drug discovery and health care products. Further and extensive research is therefore required on the safety approval and therapeutic application of the species of this genus as well as clinical trials and pharmacokinetic studies.
A new C–C linked benzophenathridine–2-quinoline dimer, and the antiplasmodial activity of alkaloids from Zanthoxylum holstzianum
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The CH₂Cl₂/MeOH (1:1) extract of Zanthoxylum holstzianum stem bark showed good antiplasmodial activity (IC₅₀ 2.5 ± 0.3 and 2.6 ± 0.3 µg/mL against the W2 and D6 strains of Plasmodium falciparum, respectively). From the extract five benzophenanthridine alkaloids [8-acetonyldihydrochelerythrine (1), nitidine (2), dihydrochelerythine (3), norchelerythrine (5), arnottianamide (8)]; a 2-quinolone alkaloid [N-methylflindersine (4)]; a lignan [4,4′-dihydroxy-3,3′-dimethoxylignan-9,9′-diyl diacetate (7)] and a dimer of a benzophenanthridine and 2-quinoline [holstzianoquinoline (6)] were isolated. The CH₂Cl₂/MeOH (1:1) extract of the root bark afforded 1, 3–6, 8, chelerythridimerine (9) and 9-demethyloxychelerythrine (10). Holstzianoquinoline (6) is new, and is the second dimer linked by a C-C bond of a benzophenanthridine and a 2-quinoline reported thus far. The compounds were identified based on spectroscopic evidence. Amongst five compounds (1-5) tested against two strains of P. falciparum, nitidine (IC₅₀ 0.11 ± 0.01 µg/mL against W2 and D6 strains) and norchelerythrine (IC₅₀ value of 0.15 ± 0.01 µg/mL against D6 strain) were the most active.
The chemosystematics of the genus Zanthoxylum L. (Rutaceae) in Kenya
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Members of this genus are also used as aphrodisiac, analgesic, febrifuge, stimulant, tonic, and homeastatic agents (Iwu, 2014; Williams, 2012). Pharmacological activities recorded on members of this genus include; antimicrobial (Kaigongi et al., 2014, 2020b; Buyinza, 2012; Matu and Van Staden 2003), anticancer (Kaigongi et al., 2020b; Özkan et al., 2013) and antimalarial (Musila et al., 2013; Nguta et al., 2010; Akampurira, 2013; Rukunga et al., 2009; Kirira et al., 2006; Were et al., 2010). Additionally, some species are used as pesticides, textile dyes and building materials (Koomson, 2003; Nussinovitch, 2009).
There is currently a significant taxonomic confusion in the genus Zanthoxylum which has a wide application in Kenyan herbal medicine. The significance of proper identification/classification of medicinal plants is key in connecting the ethnobotanical uses with the existing chemical and biological knowledge. This study aims at solving the challenges encountered in the classification of Zanthoxylum species in Kenya using metabolomics as a tool for chemosystematics.
Five Zanthoxylum species (Z. chalybeum, Z. gilletii, Z. holtzianum, Z. paracanthum and Z. usambarense) were analyzed using metabolomics approach where LC-MS analysis was used to establish the chemical profiles of each species. Hierarchical cluster analysis was used to show the relationship among the Zanthoxylum spp studied.
Based on phytochemical production, the five Zanthoxylum species segregated into two clusters; cluster one composed of Z. gilletii Z. usambarense and Z. paracanthum while Z. chalybeum and Z. holtzianum constituted cluster two. The chemical markers (discriminants) were identified as; laudanine, kalambroside A, kalambroside B, gamma-sanshool, ochotensine, cis-N-methylcanadine, 18-hydroxylinoleic acid, dihydrochelerythrine, N-feruloylglycine, p-coumaric acid ethyl ester, dihydroberberine and one unknown compound.
Chemometric analysis of relatedness of Zanthoxylum group of plants conformed to the current classical taxonomy of Zanthoxylum species in Kenya.
Medicinal plants as a fight against murine blood-stage malaria
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Malaria is an infectious parasitic disease affecting most of countries worldwide. Due to antimalarial drug resistance, researchers are seeking to find another safe efficient source for treatment of malaria. Since many years ago, medicinal plants were widely used for the treatment of several diseases. In general, most application is done first on experimental animals then human. In this article, medicinal plants as antimalarial agents in experimental animals were reviewed from January 2000 until November 2020.
In this systematic review published articles were reviewed using the electronic databases NCBI, ISI Web of knowledge, ScienceDirect and Saudi digital library to check articles and theses for M.Sc/Ph.D. The name of the medicinal plant with its taxon ID and family, the used Plasmodium species, plant part used and its extract type and the country of harvest were described.
The reviewed plants belonged to 83 families. Medicinal plants of families Asteraceae, Meliaceae Fabaceae and Lamiaceae are the most abundant for use in laboratory animal antimalarial studies. According to region, published articles from 33 different countries were reviewed. Most of malaria published articles are from Africa especially Nigeria and Ethiopia. Leaves were the most common plant part used for the experimental malaria research. In many regions, research using medicinal plants to eliminate parasites and as a defensive tool is popular.
Chemical and Cytotoxic Activity of three main Sesquiterpenoids from Warburgia ugandensis
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Citation Excerpt :
In traditional African medicine, Warburgia ugandensis is highly valued and frequently used to treat a wide range of diseases [6]. In fact, in several phytochemical studies, antibacterial [7–9], antifungal [8,9], antiparasitic [10,11] as well as antioxidant [12] and cytotoxic [9] activities were reported for W. ugandensis extracts. Interestingly, compared to other members of evergreen native tree species, W. ugandensis extracts had an oral LD50 > 5000 mg/kg body weight in mice, and no mortality was recorded in dose levels ranging from 500 to 5000 mg/kg body weight indicating safety for human use [13].
Plant materials of Warburgia ugandensis have long been used in traditional African medicine due to their broad spectrum of biological activity. Here, we describe an efficient and mild isolation of three major compounds polygodial 1a, warburganal 2a, and muzigadial 2b using Soxhlet extraction and combined preparative separation methods. Furthermore, 9-deoxymuzigadial was reported for the first time from this species. Since in silico analysis predicted drug-likeness, the compounds were tested against leukemia and solid cancer cell lines as well as human primary cells in vitro. The most susceptible cells were THP-1 indicating a selective effect against monocytes pointing towards a potential treatment of monocytic leukemia. The most robust cells were primary gingival fibroblasts offering selectivity also to solid cancers. The results in this study shed new light on long-known chemical compounds from traditional medicine and pave the way for in vivo studies to advance the substances to the next level towards the use as anti-cancer drugs.
Medicinal plants used for management of malaria among the Luhya community of Kakamega East sub-County, Kenya
2016, Journal of Ethnopharmacology
Malaria remains a major health problem worldwide especially in sub-Saharan Africa. In Kenya, 80% of the population is at risk of contracting the disease. Pregnant mothers and children under five years are the most affected by this disease. Antimalarial drug resistance poses a major threat in the fight against malaria necessitating continuous search for new antimalarial drugs. Due to inadequate and inaccessible health facilities, majority of people living in rural communities heavily depend on traditional medicine which involves the use of medicinal plants for the management of malaria. Most of these indigenous knowledge is undocumented and risks being lost yet such information could be useful in the search of new antimalarial agents.
An ethnobotanical survey was carried out among the Luhya community of Kakamega East sub-County, a malaria epidemic region, with the aim of documenting the plants used in the management of malaria.
Semi-structured questionnaires were used to collect information from 21 informants who included traditional medicine practitioners and other caregivers who had experience in use of plants in management of malaria. These were drawn from 4 villages located in Kakamega East sub-county, within Kakamega County based on their differences in topography. Information recorded included plant names, parts used, mode of preparation and administration and the sources of plant materials. A literature search was conducted using PubMed and google scholar to identify the reported traditional uses of these plants and studied antiplasmodial activities.
In this study, 57% of the informants were aged above 50 years and a total of 61% had either no formal education or had only attained primary school education. A total of 42 plant species belonging to 24 families were identified. Most plants used in the management of malaria in this community belonged to Lamiaceae (18%), Leguminosae (9%) and Compositae (9%) plant families. Plants mostly used included Melia azedarach L, Aloe spp, Ajuga integrifolia Buch. Ham, Vernonia amygdalina Del., Rotheca myricoides (Hochst.) Steane and Mabb, Fuerstia africana T.C.E.Fr., Zanthoxylum gilletii (De Wild.) P.G.Waterman and Leucas calostachys Oliv. Rumex steudelii Hochst.ex A. Rich and Phyllanthus sepialis Müll. Arg are reported for the first time in the management of malaria. Although Clerodendrum johnstonii Oliv. (Jeruto et al., 2011) and Physalis peruviana L.(Ramadan et al., 2015) are reported in other studies for management of malaria, no studies have been carried out to demonstrate their antiplasmodial activity.
The plant parts mostly used were the leaves (36%) and stem barks (26%). Majority of these plants were prepared as decoctions by boiling and allowed to cool before administration (66%) while infusions accounted for 28% of the preparations. The literature mined supports the use of these plants for the management of malaria since most of them have demonstrated in-vitro and in-vivo antiplasmodial activities.
Most of the reported plant species in this study have been investigated for antiplasmodial activity and are in agreement with the ethnomedical use. Two (2) plants are reported for the first time in the management of malaria. There is need for documentation and preservation of the rich ethnomedical knowledge within this community given that most of the practitioners are advanced in age and less educated. There is also the danger of over-exploitation of plant species as most of them are obtained from the wild, mainly Kakamega forest. Therefore, there is need for determining the economically and medicinally important plants in this community and planning for their preservation.

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Prophylactic and curative activities of extracts from Warburgia ugandensis Sprague (Canellaceae) and Zanthoxylum usambarense (Engl.) Kokwaro (Rutaceae) against Plasmodium knowlesi and Plasmodium berghei

Abstract

Ethnopharmacological relevance

Aim of study

Materials and methods

Results

Conclusion

Graphical abstract

Introduction

Section snippets

Plants

Plant extracts

In vitro activities of Warburgia ugandensis and Zanthoxylum usambarense against Plasmodium knowlesi

In vitro anti-plasmodial activities and IC50 values

Conflict of interest

Acknowledgements

Journal of Ethnopharmacology

Biochemical Pharmacology

Journal of Ethnopharmacology

Experimental Parasitology

Journal of Ethnopharmacology

Journal of Ethnopharmacology

Parasitology Today

Synergistic in vitro antimalarial activity of plant extracts used as traditional herbal remedies in Mali

Parasitology Research

Anti-plasmodial activity of seven plants used in African folk medicine

Indian Journal of Pharmacology

Plasmodium knowlesi malaria in humans is widely distributed and potentially life threatening

Clinical Infectious Diseases

Quantitative assessment of antimalarial activity in vitro by a semiautomated microdilution technique

Antimicrobial Agents Chemotherapy

Antimalarial Plants used by Hansa in Northern Nigeria

Tropical Doctor

In vitro anti-plasmodial activities and IC₅₀ values