Etudes in vitro des activités d’Artemisia annua L.

Cet article présente les études publiées concernant l’activité in vitro d’Artemisia annua L. sur les Plasmodiums, agents pathogènes à l’origine du paludisme

2005

Shi-you Li, Cong Chen, Hai-qing Zhang, Hai-yan Guo, Hui Wang, Lin Wang a,b , Xiang Zhang c , Shi-neng Hua, Jun Yu, Pei-gen Xiao, Rong-song Li, Xuehai Tan
Identification of natural compounds with antiviral activities against SARS-associated coronavirus
Antiviral Research 67 (2005) 18-23, © 2005 Elsevier B.V

Identification of natural compounds with antiviral activities against SARS-associated coronavirus

Abstract :

More than 200 Chinese medicinal herb extracts were screened for antiviral activities against Severe Acute Respiratory Syndrome-associated coronavirus (SARS-CoV) using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay for virus-induced cytopathic effect (CPE). Four of these extracts showed moderate to potent antiviral activities against SARS-CoV with 50% effective concentration (EC50) ranging from 2.4 +/- 0.2 to 88.2 +/- 7.7 microg/ml. Out of the four, Lycoris radiata was most potent. To identify the active component, L. radiata extract was subjected to further fractionation, purification, and CPE/MTS assays. This process led to the identification of a single substance lycorine as an anti-SARS-CoV component with an EC50 value of 15.7 +/- 1.2 nM. This compound has a CC50 value of 14980.0 +/- 912.0 nM in cytotoxicity assay and a selective index (SI) greater than 900. The results suggested that four herbal extracts and the compound lycorine are candidates for the development of new anti-SARS-CoV drugs in the treatment of SARS.

2009

Taísa Carrijode Oliveira, Deise A. Oliveira Silva, Cristina Rostkowska, Samantha Ribeiro Béla, Eloisa A.V.Ferro, Pedro Mellilo Magalhães, José Roberto Mineo
Toxoplasma gondii : Effects of Artemisia annua L. on susceptibility to infection in experimental models in vitro and in vivo
Experimental Parasitology, Volume 122, Issue 3, July 2009, Pages 233-241

Toxoplasma gondii : Effects of Artemisia annua L. on susceptibility to infection in experimental models in vitro and in vivo

Abstract :

Considering that the treatment for toxoplasmosis is based on drugs that show limited efficacy due to their substantial side effects, the purpose of the present study was to evaluate the effects of Artemisia annua on in vitro and in vivo Toxoplasma gondii infection. A. annua infusion was prepared from dried herb and tested in human foreskin fibroblasts (HFF) or mice that were infected with the parasite and compared with sulfadiazine treatment. For in vitro experiments, treatment was done on parasite before HFF infection or on cells previously infected with T. gondii and the inhibitory concentration (IC50) values for each treatment condition were determined. Viability of HFF cells in the presence of different concentrations of A. annua infusion and sulfadiazine was above 72%, even when the highest concentrations from both treatments were tested. Also, the treatment of T. gondii tachyzoites with A. annua infusion before infection in HFF cells showed a dose–response inhibitory curve that reached up to 75% of inhibition, similarly to the results observed when parasites were treated with sulfadiazine. In vivo experiments with a cystogenic T. gondii strain demonstrated an effective control of infection using A. annua infusion. In conclusion, our results indicate that A. annua infusion is useful to control T. gondii infection, due to its low toxicity and its inhibitory action directly against the parasite, resulting in a well tolerated therapeutic tool.

2010

Gueye Papa El hadji Omar
Chimiosensibilité in vitro de Plasmodium falciparum par rapport à la tisane d’Artemisia annua et action stérilisante de la même tisane sur les eaux polluées
Thèse de doctorat en Biologie et Pathologie Humaine (ED-SEV) Octobre 2010, Hôpital Aristide Le Dantec, Laboratoire Bactériologie Virologie, Section Malaria, Dakar Sénégal

Full text submitted to request

Discussion et conclusions :

Dans l’étude présentée ici, nous avons étudié la chimiosensibilité in vitro des souches de Plasmodium falciparum par rapport à la tisane d’Artemisia annua en provenance de Luxembourg. Cette tisane est constituée de 0,2% d’artémisinine. La méthode d’extraction utilisée est celle décrite par Rath et al, 2004 selon laquelle 70%1 de cette teneur en artémisinine seraient extraits après infusion. Notre but consistait à étudier l’effet de la tisane sur les différentes souches de terrains obtenues dans le poste de Santé de Touba Diacksao mais aussi sur des souches de références que sont les 3D7 et W2. Les tests ont montré des valeurs de CI50 qui varient avec un maximum à 3,545 µmol/L et un minimum à 0,087µmol/l. La moyenne est de 0,494 µmol/l ou 494 nM/L. Ces valeurs comparées à celles trouvées dans la littérature pour l’artémisinine pure et de ses dérivés seraient nettement supérieures.

Par contre, si nous cherchons dans la littérature des valeurs de référence pour notre CI50 tel que exprimé 0,139 µg/ml, nous trouvons une valeur fort acceptable. Une étude tanzanienne menée par Malebo HM et al, 2009 a montré que les fortes activités antiplasmodiales, de la tisane d’Artemisia annua et d’autres plantes médicinales étaient observées pour des CI50 inférieures à 0,5 µg/ml. Ceci nous permettrait de dire que notre tisane luxembourgeoise aurait une forte activité antiplasmodiale, et cela bien qu’elle n’est pas aussi riche en artémisinine que d’autres tisanes comme par exemple celle de Brésil où il y a une teneur de 1% en artémisinine. Ceci laisserait penser que cette forte activité antipaludique de la tisane luxembourgeoise serait due, non seulement à l’artémisinine seule, mais à l’effet combiné d’autres substances qui s’y trouvent (coumarine et scopolétine, polyphénols, flavonoides …). Ceci est quelque peu confirmé par une étude éthiopienne où des effets thérapeutiques antipaludiques ont été observés avec une tisane ne contenant que 0.01% d’artémisinine (A Muzemil, PhD thesis, Addis Ababa 2008).

Conclusion et perspective :

En conclusion les tests in vitro sur les souches de Plasmodium falciparum montrent que la tisane luxembourgeoise serait efficace. Cette efficacité démontrée in vitro contre Plasmodium falciparum, malgré une teneur en artémisinine faible, nécessite des études plus approfondies qui vont surtout porter sur la concentration des autres substances (polyphénols, camphre, coumarine....) présentes dans la tisane. Ceci permettrait éventuellement de mieux comprendre les différences trouvées pour nos CI50, si on les compare, soit aux CI50 de l’artemisinine pure, soit aux CI50 d’autres tisanes. Il est fort probable qu’il faut garder une grande prudence à vouloir extrapoler les valeurs obtenues in vitro pour des substances pures à l’effet in vivo. Les travaux de Pedro Melillo de Magalhaes à l’Université de Louvain comme ceux antérieurement de Karin Raeth ont claiement montré une nette différence pour la pharmacocinétique entre substance pure et tisane.
D’autres pistes de recherche sont envisagées également comme : l’effet de la tisane sur le systême immunitaire, sur les lymphocytes, les cellules cancéreuses. Nous comptons aussi continuer cette étude avec des tisanes d’Artemisia annua d’autres origines bien caractérisées par les analyses de Rosine Chougouo, docteur en pharmacie de l’Université des Montagnes du Cameroun.

Étude sur les eaux de pluie :

Dans le cadre de l’étude de l’efficacité de la tisane sur les bactéries, nous avons mené une campagne aux mois d’Août et Septembre dans 12 villages de la zone rurale de Rufisque (Dakar-Sénégal). Cette campagne consistait à déterminer la qualité bactériologique des eaux de puits avant et après traitement avec la tisane d’Artemisia annua.

L’évaluation bactériologique de l’eau a été faite à partir d’une méthode d’analyse utilisant une mallette Protatest Waggtech qui permettait de dénombrer, en présence d’auryl sulfate comme milieu de culture, la présence de coliformes fécaux. La qualité physique de l’eau a aussi été déterminée en utilisant les paramètres comme le pH, la température, la conductivité et la turbidité. Les puits prélevés proviennent de 12 villages et concernent les puits les plus fréquentés dans chaque localité. Sur un total de 12 puits, 10 ont été contaminés parfois à des degrés très avancés par des coliformes fécaux.

Conclusion :

Après incubation avec la tisane d’Artemisia annua, nous avons observé une élimination totale des coliformes fécaux après 14h et 20h d’incubation. Ces résultats obtenus montrent que la tisane d’Artemisia annua est efficace sur les coliformes fécaux. Ce travail fera l’objet de publication.

2012

Luiz Francisco Rocha e Silva, Pedro Melillo de Magalhães, Mônica Regina Farias Costa, Maria das Graças Costa Alecrim, Francisco Célio Maia Chaves, Ari de Freitas Hidalgo, Adrian Martin Pohlit, Pedro Paulo Ribeiro Vieira
In vitro susceptibility of Plasmodium falciparum Welch field isolates to infusions prepared from Artemisia annua L. cultivated in the Brazilian Amazon
Mem. Inst. Oswaldo Cruz, vol. 107 no. 7 Rio de Janeiro November 2012

In vitro susceptibility of Plasmodium falciparum Welch field isolates to infusions prepared from {Artemisia annua} L. cultivated in the Brazilian Amazon

Abstract :

Artemisinin is the active antimalarial compound obtained from the leaves of Artemisia annua L. Artemisinin, and its semi-synthetic derivatives, are the main drugs used to treat multi-drug-resistant Plasmodium falciparum (one of the human malaria parasite species). The in vitro susceptibility of Plasmodium falciparum K1 and 3d7 strains and field isolates from the state of Amazonas, Brazil, to Artemisia annua infusions (5 g dry leaves in 1 L of boiling water) and the drug standards chloroquine, quinine and artemisinin were evaluated. The Artemisia annua used was cultivated in three Amazon ecosystems (várzea, terra preta de índio and terra firme) and in the city of Paulínia, state of São Paulo, Brazil. Artemisinin levels in the Artemisia annua leaves used were 0.90-1.13% (m/m). The concentration of artemisinin in the infusions was 40-46 mg/L. Field Plasmodium falciparum isolates were resistant to chloroquine and sensitive to quinine and artemisinin. The average 50% inhibition concentration values for Artemisia annua infusions against field isolates were 0.11-0.14 μL/mL (these infusions exhibited artemisinin concentrations of 4.7-5.6 ng/mL) and were active in vitro against Plasmodium falciparum due to their artemisinin concentration. No synergistic effect was observed for artemisinin in the infusions. Key words : Artemisia annua L. - infusion - antiplasmodial - in vitro - field isolate - Amazon Region

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Antonella De Donno, Adele Idolo, Marcello Guido, Alessandro Caccioppola, Luciano Villanova, Alessandro Merendino, Francesco Bagordo, Francesco Paolo Fanizzi
First-time comparison of the in vitro antimalarial activity of Artemisia annua herbal tea and artemisinin
Transactions of the Royal Society of Tropical Medicine and Hygiene 106(11) : 696-700 · September 2012

First-time comparison of the in vitro antimalarial activity of {Artemisia annua} herbal tea and artemisinin

Abstract :

Artemisia annua tea has been proven to be a very effective treatment for malaria in various clinical trials, but to date its efficacy has not been investigated in vitro. A study was therefore performed to evaluate the effects of Artemisia annua tea on Plasmodium falciparum cultures in vitro. The concentration of artemisinin in the herbal tea preparation was also determined. The herbal tea extract was tested against chloroquine (CQ)-sensitive D10 and CQ-resistant W2 strains of Plasmodium falciparum using the parasite lactate dehydrogenase assay. Quantification of artemisinin in the extract of leaves of Artemisia annua was performed using proton nuclear magnetic resonance (1H-NMR). Results of the in vitro tests were consistent with the clinical efficacy of Artemisia annua tea [50% inhibitory concentration (IC50) for strain D10 = 1.11 ± 0.21 μg/ml ; IC50 for strain W2 = 0.88 ± 0.35 μg/ml]. The concentration of artemisinin in Artemisia annua tea (0.18 ± 0.02% of dry weight) was far too low to be responsible for the antimalarial activity. The artemisinin present in the tea is probably co-solubilised with other ingredients, some of which also have antimalarial activity and act synergistically with it. These compounds also merit further research to determine whether their presence hinders the development of parasite resistance compared with pure artemisinin.
Résumé : Le thé d’Artemisia annua s’est avéré être un traitement très efficace contre le paludisme lors de divers essais cliniques, mais son efficacité n’a pas encore été étudiée in vitro. Une étude a donc été réalisée pour évaluer les effets du thé Artemisia annua sur des cultures de Plasmodium falciparum in vitro. La concentration d’artémisinine dans la préparation de la tisane a également été déterminée. L’extrait de tisane a été testé contre les souches D10 sensibles à la chloroquine (CQ) et W2 résistantes à la CQ de Plasmodium falciparum en utilisant le test de la lactate déshydrogénase du parasite. La quantification de l’artémisinine dans l’extrait de feuilles d’Artemisia annua a été effectuée par résonance magnétique nucléaire du proton (RMN 1H). Les résultats des tests in vitro étaient conformes à l’efficacité clinique du thé Artemisia annua [concentration inhibitrice de 50 % (CI50) pour la souche D10 = 1,11 ± 0,21 μg/ml ; CI50 pour la souche W2 = 0,88 ± 0,35 μg/ml]. La concentration d’artémisinine dans le thé Artemisia annua (0,18 ± 0,02 % du poids sec) était bien trop faible pour être responsable de l’activité antipaludique. L’artémisinine présente dans le thé est probablement co-solubilisée avec d’autres ingrédients, dont certains ont également une activité antipaludique et agissent en synergie avec elle. Ces composés méritent également des recherches plus approfondies afin de déterminer si leur présence entrave le développement de la résistance des parasites par rapport à l’artémisinine pure.

***

Andrea Lubbe, Isabell Seiber, Thomas Klimkait, Frank van der Kooy
Ethnopharmacology in overdrive : The remarkable anti-HIV activity of Artemisia annua
Journal of Ethnopharmacology, 2012 June 14 ; 141 (3) : 854-9.
Ethnopharmacology in overdrive : The remarkable anti-HIV activity of Artemisia annua Journal of Ethnopharmacology. Elsevier

Full text submitted to request

Abstract :

Ethnopharmacological relevance : Artemisia annua contains the well-known antimalarial compound artemisinin, which forms the backbone of the global malaria treatment regime. In African countries a tea infusion prepared from Artemisia annua has been used for the treatment of malaria only for the past 10-20 years. Several informal claims in Africa exist that the Artemisia annua tea infusions are also able to inhibit HIV. Since HIV is a relatively newly emerged disease, the claims, if substantiated, could provide a very good example of "ethnopharmacology in overdrive". The objective of this study was to provide quantitative scientific evidence that the Artemisia annua tea infusion exhibits anti-HIV activity through in vitro studies. A second objective was to determine if artemisinin plays a direct or indirect (synergistic) role in any observed activity. This was done by the inclusion of a chemically closely related species, Artemisia afra, known not to contain any artemisinin in our studies.

Materials and methods : Validated cellular systems were used to test Artemisia annua tea samples for anti-HIV activity. Two independent tests with different formats (an infection format and a co-cultivation format) were used. Samples were also tested for cellular toxicity against the human cells used in the assays.
Results : The Artemisia annua tea infusion was found to be highly active with IC(50) values as low as 2.0 μg/mL. Moreover we found that artemisinin was inactive at 25 μg/mL and that a chemically related species Artemisia afra (not containing artemisinin) showed a similar level of activity. This indicates that the role of artemisinin, directly or indirectly (synergism), in the observed activity is rather limited. Additionally, no cellular toxicity was seen for the tea infusion at the highest concentrations tested.

Conclusion : This study provides the first in vitro evidence of anti-HIV activity of the Artemisia annua tea infusion. We also report for the first time on the anti-HIV activity of Artemisia afra although this was not an objective of this study. These results open the way to identify new active pharmaceutical ingredients in Artemisia annua and thereby potentially reduce the cost for the production of the important antimalarial compound artemisinin.

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Andrea Lubbe, Isabell Seiber, Thomas Klimkait, Frank van der Kooy
Ethnopharmacology in overdrive : The remarkable anti-HIV activity of Artemisia annua
Journal of Ethnopharmacology, 2012 June 14 ; 141 (3) : 854-9.

Abstract :

Ethnopharmacological relevance : Artemisia annua contains the well-known antimalarial compound artemisinin, which forms the backbone of the global malaria treatment regime. In African countries a tea infusion prepared from Artemisia annua has been used for the treatment of malaria only for the past 10-20 years. Several informal claims in Africa exist that the Artemisia annua tea infusions are also able to inhibit HIV. Since HIV is a relatively newly emerged disease, the claims, if substantiated, could provide a very good example of "ethnopharmacology in overdrive". The objective of this study was to provide quantitative scientific evidence that the Artemisia annua tea infusion exhibits anti-HIV activity through in vitro studies. A second objective was to determine if artemisinin plays a direct or indirect (synergistic) role in any observed activity. This was done by the inclusion of a chemically closely related species, Artemisia afra, known not to contain any artemisinin in our studies.

Materials and methods : Validated cellular systems were used to test Artemisia annua tea samples for anti-HIV activity. Two independent tests with different formats (an infection format and a co-cultivation format) were used. Samples were also tested for cellular toxicity against the human cells used in the assays.

Results : The Artemisia annua tea infusion was found to be highly active with IC(50) values as low as 2.0 μg/mL. Moreover we found that artemisinin was inactive at 25 μg/mL and that a chemically related species Artemisia afra (not containing artemisinin) showed a similar level of activity. This indicates that the role of artemisinin, directly or indirectly (synergism), in the observed activity is rather limited. Additionally, no cellular toxicity was seen for the tea infusion at the highest concentrations tested.

Conclusion : This study provides the first in vitro evidence of anti-HIV activity of the Artemisia annua tea infusion. We also report for the first time on the anti-HIV activity of Artemisia afra although this was not an objective of this study. These results open the way to identify new active pharmaceutical ingredients in Artemisia annua and thereby potentially reduce the cost for the production of the important antimalarial compound artemisinin.

2013

Julia Mouton, Olivia jansen, Michel Frédérich, Frank van der Koyy
Is Artemisinin the Only Antiplasmodial Compound in the Artemisia annua Tea Infusion ? An in Vitro Study
Planta medica 2013 Apr ;79(6):468-70. doi : 10.1055/s-0032-1328324. Epub 2013 Mar 19.

Abstract : In our ongoing investigation into Artemisia annua for the treatment of malaria, we decided to study the possibility that synergism might enhance the efficacy of artemisinin. Our main objective was to test tea infusions and nonpolar extracts prepared from different Artemisia annua varieties against Plasmodium falciparum in vitro in order to determine if synergism will increase the effectiveness of artemisinin in the samples as compared to pure artemisinin. We found that the IC50 of artemisinin in the tea and nonpolar extracts was not significantly different to the IC50 of pure artemisinin. We could show that the year and country of harvest or storage conditions did not have any influence on the activity and that it narrowly followed the concentration of artemisinin in all the extracts. In conclusion, based on these in vitro results, artemisinin seems to be the only active antiplasmodial compound in Artemisia annua.

Gueye Papa El hadji Omar, Diallo Mouhamadou, Deme Awa Bineta, Badiane Aida Sadikh, Dior Diop Mare1, Ahouidi Ambroise, Abdoul Aziz Ndiaye, Dieng Thérése, Lutgen Pierre, Mboup Souleymane and Sarr Ousmane
Tea Artemisia annua inhibits Plasmodium falciparum isolates collected in Pikine, Senegal
African Journal of Biochemistry Research, Vol. 7(7), pp. 107-1 13, August 2013

Tea A annua inhibits Plasmodium falciparum isolates collected in Pikine, Senegal

Abstract :

Malaria is a major scourge of most countries in Africa which continues to defy science and technology. Several medicinal plants are traditionally used for the treatment of malaria and other protozoa infections. We aimed to assess by the Double-site Enzyme-linked Lactate dehydrogenase enzyme Immuno-detection (DELI) test for the first time in Senegal, Plasmodium falciparum isolates in vitro susceptibility to Tea Artemisia annua (TAA). In total, 40 field isolates have been tested and the mean IC50 was 0.095 µg/ml, while the IC50 for the 3D7 and W2 laboratory adapted strains were 0.14 and 0.39 µg/ml, respectively. Tea Artemisia annua sensitivity was not obtained for three isolates because of lack of growth. The results suggest that Artemisia annua tea has potent antiplasmodial activity against Plasmodium falciparum strains collected in Pikine, Senegal.

Key words : Artemisia annua, enzyme-linked lactate dehydrogenase enzyme immuno-detection (DELI), Plasmodium falciparum, IC50.

2014

Papa Elhadji Omar Gueye, Baba Dièye, Mouhamadou Diallo, Amy Kristine Bei, Aminata Mbaye, Souleymane Mboup, Pierre Lutgen, Dyann F. Wirth, Daouda Ndiaye
DAPI - test for evaluation ex vivo susceptibility on Plasmodium falciparum isolates collected in Thiès, Senegal to Artemisia annuaL.

Abstract :

In Senegal, where malaria is endemic throughout the country, there is clear evidence that many people who need to be treated with this combination of antimalarials do not have access to it. This situation results primarily from poverty. Therefore, it is necessary to define new strategies to alleviate the burden represented by the infection ; it is also urgent to develop new ways and means to make it possible for all Senegalese citizens to afford to pay for the available treatment. One way to reach this goal is through the use of the herbal tea of Artemisia annua. We aimed at assessing by the Di Amidino Phenyl Indol (DAPI) test for the first time in Senegal, Plasmodium falciparum isolates ex vivo susceptibility to Tea Artemisia annua (TAA) from Luxembourg, relatively poor in artemisinine (0.2%).

Overall, 45 fresh samples collected from the study participants were tested and 42 Plasmodium falciparum isolates adapted to the ex vivo growth. The geometric average IC50 was 1.034 μg/ml. The IC50 values were lower than 5 μg/ml for 41 samples and only one exhibited an IC50 > 5 μg/ml. The IC50 value was lower than 0.5 μg/ml for 16 samples, 25 samples had IC50 ranging from 0.5μg/ml to 5μg/ml and only one sample had IC50 from 5μg/ml to 10μg/ml. The IC50 values were undetermined for 3 isolates for unknown reasons. The 3D7 and W2 laboratory adapted strains respectively exhibited IC50 of 1.040 and 1.394 μg/ml. The results suggest that TAA has a potent anti-plasmodial activity against Plasmodium falciparum strains collected in Thiès, Senegal.

Keys words : Artemisia annua, Di Amidino Phenyl Indol (DAPI), Plasmodium falciparum.

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2014

Mohammad Islamuddin, Garima Chouhan, Muzamil Y. Want, Maujiram Tyagi, Malik Z. Abdin, Dinkar Sahal, and Farhat Afrin
Leishmanicidal activities of Artemisia annua leaf essential oil against Visceral Leishmaniasis
Frontiers in Microbiology. 2014 ; 5 : 626

Leishmanicidal activities of Artemisia annua leaf essential oil against Visceral Leishmaniasis

Abstract :

Visceral leishmaniasis (VL), the second-most dreaded parasitic disease after malaria, is currently endemic in 88 countries. Dramatic increases in the rates of infection, drug resistance, and non-availability of safe vaccines have highlighted the need for identification of novel and inexpensive anti-leishmanial agents from natural sources. In this study, we showed the leishmanicidal effect of essential oil from Artemisia annua leaves (AALEO) against Leishmania donovani in vitro and in vivo. AALEO was extracted by hydrodistillation and characterized by GC-MS, the most abundant compounds were found to be camphor (52.06 %) followed by β-caryophyllene (10.95 %). AALEO exhibited significant leishmanicidal activity against L. donovani, with 50 % inhibitory concentration of 14.63 ± 1.49 μg ml-1 and 7.3 ± 1.85 μg ml-1, respectively, against the promastigotes and intracellular amastigotes. The effect was mediated through programmed cell death as confirmed by externalization of phosphatidylserine, DNA nicking by TdT-mediated dUTP nick-end labeling assay, dyskinetoplastidy, cell cycle arrest at sub-G0–G1 phase, loss of mitochondrial membrane potential and reactive oxygen species generation in promastigotes and nitric oxide generation in ex vivo model. AALEO presented no cytotoxic effects against mammalian macrophages even at 200 μg ml-1. Intra-peritoneal administration of AALEO (200 mg/ kg.b.w.) to infected BALB/c mice reduced the parasite burden by almost 90% in the liver and spleen with significant reduction in weight. There was no hepato- or nephro-toxicity as demonstrated by normal levels of serum enzymes. The promising antileishmanial activity shown by camphor-rich AALEO may provide a new lead in the treatment of VL.

Keywords : leishmaniasis, visceral, essential oil, Artemisia annua, leishmanicidal, apoptosis, therapeutic efficacy

2015

Hermine Zime-Diawara, Habib Ganfon, Fernand Gbaguidi, Achille Yemoa, Joanne Bero, Olivia Jansen, Brigitte Evrard, Mansourou Moudachirou, Michel., Frédérich, Joëlle Quetin-Leclercq
Antimalarial action of aqueous and hydro alcoholic extracts of Artemisia annua L . cultivated in Benin : In vitro and in vivo studies
Journal of Chemical and Pharmaceutical Research, 2015, 7(8):817-823

Antimalarial action of aqueous and hydro alcoholic extracts of {Artemisia annua} L . cultivated in Benin : In vitro and in vivo studies

Abstract : The aqueous and hydro alcoholic extracts of Artemisia annua plants acclimatized and set in culture in Benin, were investigated for their in vitro and in vivo antimalarial value. These extracts were prepared by decoction and maceration using water and ethanol, and were tested for in vitro activity against the strains of Plasmodium falciparum. Those extracts were also investigated in vivo against Plasmodium berghei NK 173 infected mice by the 4-day suppressive test. The action of these extracts was similar in vitro to the action of pure artemisinin at the same dosage. But, the in vivo studies on mice show that we have the same efficacy with aqueous extract of Artemisia annua (artemisinin content of 20 mg/kg) than with pure artemisinin at a dosage of 140 mg/kg. The hydro alcoholic extract of Artemisia annua (artemisinin content of 20 mg/kg) was better than the both other conditions. The others components of the plant Artemisia annua potentiate artemisinin activity on Plasmodium. The aqueous and hydro alcoholic extracts of Artemisia annua L. are very effective on malaria and they can be used by populations to cure this disease, especially in endemic areas where modern medicines are expensive.
Keywords :Artemisia annua extracts, in vitro studies, in vivo studies, artemisinin potentiation.

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Amivi Tété-Bénissan, Mlatovi Dégbé, Françoise Debierre-Grockiego, Isabelle Dimier-Poisson, Kodjo Aklikokou, Messanvi Gbeassor
Etude préliminaire Sur l’effet des extraits de feuilles d’Artemisia annua et de Moringa oleifera sur Toxoplasmagondii in vitro
Actes des 16 èmes Journées Scientifiques Annuelles de la SOACHIM ; 03 – 06 Août 2015 ; Abidjan – Côte d’Ivoire

Etude préliminaire Sur l’effet des extraits de feuilles d’Artemisia annua et de Moringa oleifera sur Toxoplasmagondii in vitro

Résumé : La toxoplasmose est causée par Toxoplasma gondii, un protozoaire intracellulaire obligatoire. Sa séroprévalence varie de 50 à 70% dans les régions humides de l’Afrique comme le Togo. L’infection est bénigne chez le sujet immunocompétent mais entraîne des conséquences graves chez la femme enceinte et les sujets immunodéprimés. Actuellement, le traitement de première intention est l’association pyriméthamine-sulfamide (molécules toxiques, tératogènes avec des résistances parasitaires). Dans le cadre de la recherche de traitement alternatif, les effets toxoplasmicides des extraits alcooliques et hydro alcooliques des feuilles d’Artemisia annua et de Moringa oleifera sont testés in vitro sur Toxoplasma gondii et les cellules HFF. Les résultats indiquent que les deux extraits d’Artemisia annua ont un effet inhibiteur sur Toxoplasma gondii avec des CI 50 respectives de 1.18µg/ml et de 9.50µg/ml. Les tests de cytotoxicité révèlent que l’extrait hydo alcoolique d’Artemisia annua est plus toxique que l’extrait alcoolique. Les extraits de Moringa oleifera ne présentent aucune toxicité vis–à-vis des cultures de cellules HFF. Les extraits alcooliques d’Artemisia annua et de Moringa oleifera ont inhibé de l’invasion des cellules in vitro par Toxoplasma gondii. Ces résultats suggèrent qu’Artemisia annua serait un bon candidat pour les recherches sur les phytomédicaments à activité toxoplasmicide
Mots Clés : Toxoplasma gondii ; Artemisia annua ; Moringa oleifera ; effets in vitro.

Abstract : Toxoplasmosis is an infection caused by the obligate intracellular parasite Toxoplasma gondii. Its seroprevalence varied from 50 to 70% in the wet areas of Africa as Togo. The infection is benign in immunocompetent subjects but involved serious consequences in the immunocompromised patients or during pregnancy. Sulfadiazine plus pyrimethamine are currently, the drugs of first choice for treatment of toxoplasmosis. Although this therapy is frequently associated with many side effects and not well tolerated. Thus, to find out less toxic drug active against the parasite, the purpose of this present study was to evaluate, the effects of Artemisia annua and Moringa oleifera alcoholic and hydro alcoholic extracts on vitro Toxoplasma gondii and HF cells. The results indicate that Artemisia annua extracts have an inhibiting effect on Toxoplasma gondii with respective IC 50, 1.18µg/ml and 9.50µg/ml. Cytotoxicity tests reveal that hydo alcoholic extract of Artemisia annua is more toxic than alcoholic extract. Moringa oleifera extracts do not have any toxicity on HFF cells. The alcoholic extracts of Artemisia annua and Moringa oleifera inhibited invasion of in vitro cells by Toxoplasma gondii. These results suggest that Artemisia annua is a good candidate for research on the drugs with toxoplasmicide activity.

Keywords : Toxoplasma gondii ; Artemisia annua ; Moringa oleifera ; in vitro effect.

2020

Cao R, Hu H, Li Y, Wang X, Xu M, Liu J, et al.
Anti-SARS-CoV-2 potential of artemisinins in vitro
ACS Infect Dis 2020 ; 6(9) : 2524–31.

Abstract

The discovery of novel drug candidates with anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) potential is critical for the control of the global COVID-19 pandemic. Artemisinin, an old antimalarial drug derived from Chinese herbs, has saved millions of lives. Artemisinins are a cluster of artemisinin-related drugs developed for the treatment of malaria and have been reported to have multiple pharmacological activities, including anticancer, antiviral, and immune modulation. Considering the reported broad-spectrum antiviral potential of artemisinins, researchers are interested in whether they could be used to combat COVID-19. We systematically evaluated the anti-SARS-CoV-2 activities of nine artemisinin-related compounds in vitro and carried out a time-of-drug-addition assay to explore their antiviral mode of action. Finally, a pharmacokinetic prediction model was established to predict the therapeutic potential of selected compounds against COVID-19. Arteannuin B showed the highest anti-SARS-CoV-2 potential with an EC50 of 10.28 ± 1.12 μM. Artesunate and dihydroartemisinin showed similar EC50 values of 12.98 ± 5.30 μM and 13.31 ± 1.24 μM, respectively, which could be clinically achieved in plasma after intravenous administration. Interestingly, although an EC50 of 23.17 ± 3.22 μM was not prominent among the tested compounds, lumefantrine showed therapeutic promise due to high plasma and lung drug concentrations after multiple dosing. Further mode of action analysis revealed that arteannuin B and lumefantrine acted at the post-entry step of SARS-CoV-2 infection. This research highlights the anti-SARS-CoV-2 potential of artemisinins and provides leading candidates for anti-SARS-CoV-2 drug research and development.

Keywords : COVID-19 ; SARS-CoV-2 ; antiviral drug ; artemisinin ; drug repurposing.

2021

Kutub Ashraf, Shahin Tajeri1, Christophe-Sébastien Arnold, Nadia Amanzougaghene, Jean-François Franetich, Amélie Vantaux, Valérie Soulard, Mallaury Bordessoulles, Guillaume Cazals, Teun Bousema, Geert-Jan van Gemert, Roger Le Grand, Nathalie Dereuddre-Bosquet, Jean-Christophe Barale, Benoit Witkowski, Georges Snounou, Romain Duval, Cyrille Y. Botté, Dominique Mazier
Artemisinin-1 independent inhibitory activity of Artemisia sp. infusions against different Plasmodium stages including relapse causing hypnozoites
bioRxiv preprint doi : https://doi.org/10.1101/2021.08.10.455849 ; this version posted August 11, 2021

Les thérapies combinées à base d’artémisinine (ACT) sont les traitements de première ligne contre le paludisme dans le monde entier. Récemment, l’utilisation d’infusions traditionnelles d’Artemisia annua (à partir de laquelle l’artémisinine est obtenue) ou d’Artemisia afra (dépourvue d’artémisinine) a été préconisée de manière controversée. Ces remèdes non réglementés à base de plantes sont fortement déconseillés car ils pourraient constituer des thérapies sous-optimales et favoriser la résistance aux médicaments. Nous avons mené la première étude comparative des effets antipaludiques des deux infusions de plantes in vitro contre les stades érythrocytaires asexués de P. falciparum et les stades pré-érythrocytaires (c’est-à-dire le foie) de diverses espèces de Plasmodium. De faibles concentrations de l’une ou l’autre des infusions ont permis d’obtenir des activités inhibitrices significatives pour chaque espèce de parasite et chaque stade étudié. Nous montrons que ces effets antiplasmodiaux étaient essentiellement indépendants de l’artémisinine et qu’ils étaient également contrôlés par des observations de l’apicoplaste et de la mitochondrie du parasite. En particulier, les perfusions ont considérablement neutralisé les sporozoïtes et, pour Plasmodium vivax et Plasmodium cynomolgi, perturbé les hypnozoïtes. Ceci constitue la première indication que des composés autres que les 8- aminoquinolines pourraient être des antipaludiques efficaces contre les parasites récidivants. Ces observations plaident en faveur d’un criblage plus approfondi afin de découvrir de nouveaux composés antipaludiques dont le besoin est urgent.
Abstract

Artemisinin-based combination therapies (ACT) are the frontline treatments against malaria worldwide. Recently the use of traditional infusions from Artemisia annua (from which artemisinin is obtained) or A. afra (lacking artemisinin) has been controversially advocated. Such unregulated plant-based remedies are strongly discouraged as they might constitute sub-optimal therapies and promote drug resistance. Here, we conducted the first comparative study of the anti-malarial effects of both plant infusions in vitro against the asexual erythrocytic stages of P. falciparum and the pre-erythrocytic (i. e., liver) stages of various Plasmodium species. Low concentrations of either infusion accounted for significant inhibitory activities across every parasite species and stage studied. We show that these antiplasmodial effects were essentially artemisinin-independent and were additionally monitored by observations of the parasite apicoplast and mitochondrion. In particular, the infusions significantly incapacitated sporozoites, and for P. vivax and P. cynomolgi, disrupted the hypnozoites. This provides the first indication that compounds other than 8- aminoquinolines could be effective antimalarials against relapsing parasites. These observations advocate for further screening to uncover urgently needed novel antimalarial lead compounds.

Gruessner BM, Weathers PJ. In vitro analyses of Artemisia extracts on Plasmodium falciparum suggest a complex antimalarial effect. PLoS One. 2021 Mar 2 ;16(3):e0240874. doi : 10.1371/journal.pone.0240874. ECollection 2021. PMID : 33651845 Free PMC article.

Le traitement antipaludique à base de feuilles séchées d’Artemisia annua L. s’est avéré efficace lors d’études antérieures sur l’animal et l’homme, mais on sait peu de choses sur son mécanisme d’action. Nous avons utilisé ici les Concentration inhibitrice médiane (IC50) et les essais en phase de cycle pour comparer les extraits d’Artemisia annua (DLAe) à l’artémisinine (ART) et à ses dérivés dans leur capacité à inhiber et à tuer les souches 3D7, MRA1252, MRA1240, Cam3.11 et Cam3.11rev de Plasmodium falciparum in vitro. Les souches ont été synchronisées par sorbitol et Percoll pour enrichir les parasites au stade de l’anneau qui ont été traités avec des extraits d’eau chaude, de méthanol et de dichlorométhane de DLA, d’artémisinine, de CoArtem™ et de dihydroartémisinine.
Des extraits d’Artemisia afra SEN ont également été testés. Il y avait une corrélation entre la concentration de l’artémisine et l’inhibition de la croissance du parasite. Bien qu’à 6 heures d’incubation du médicament, les essais en phase de cycle pour Cam3.11rev aient montré que les feuilles séchées d’Artemisia annua et l’artémisine étaient moins efficaces que CoArtem™ à forte dose, les incubations de 8 et 24 heures ont donné des résultats antiparasitaires équivalents.
Pour Cam3.11, la durée d’incubation du médicament n’a eu aucun effet. L’extrait à base de feuilles séchées d’Artemisia annua a été plus efficace sur la souche résistante MRA-1240 que sur la souche sensible MRA-1252. Comme les résultats n’étaient pas aussi robustes que ceux observés dans les études animales et humaines, une interaction avec l’hôte a été suspectée, de sorte que des sérums recueillis auprès de patients adultes et pédiatriques atteints de paludisme au Kenya ont été utilisés dans des expériences d’inhibition de l’essai en phase de cycle et comparés à des sérums d’adultes naïfs de la maladie.
Les sérums des patients paludéens des deux groupes d’âge ont inhibé la croissance parasitaire ≥ 70 % après traitement par extraits à base de feuilles séchées d’Artemisia annua et par rapport aux sujets naïfs de paludisme, ce qui suggère une certaine interaction de l’hôte avec les feuilles séchées d’Artemisia annua.
La divergence entre ces données et les rapports in-vivo suggère que les effets des feuilles séchées d’Artemisia annua nécessitent une interaction avec l’hôte pour libérer leur potentiel en tant que thérapie antipaludique. Bien que nous ayons montré qu’il existe des effets sur l’hôte basés sur le sérum qui peuvent tuer jusqu’à 95 % des parasites in vitro, on ne sait toujours pas comment ni si ces effets jouent un rôle in vivo. Ces résultats nous permettent de mieux comprendre comment l’extrait d’Artemisia annua agit contre le parasite du paludisme in vitro.

Résumé publié

Dried-leaf Artemisia annua L. (DLA) antimalarial therapy was shown effective in prior animal and human studies, but little is known about its mechanism of action. Here IC50s and ring-stage assays (RSAs) were used to compare extracts of A. annua (DLAe) to artemisinin (ART) and its derivatives in their ability to inhibit and kill Plasmodium falciparum strains 3D7, MRA1252, MRA1240, Cam3.11 and Cam3.11rev in vitro. Strains were sorbitol and Percoll synchronized to enrich for ring-stage parasites that were treated with hot water, methanol and dichloromethane extracts of DLA, artemisinin, CoArtem™, and dihydroartemisinin. Extracts of A. afra SEN were also tested. There was a correlation between ART concentration and inhibition of parasite growth. Although at 6 hr drug incubation, the RSAs for Cam3.11rev showed DLA and ART were less effective than high dose CoArtem™, 8 and 24 hr incubations yielded equivalent antiparasitic results. For Cam3.11, drug incubation time had no effect. DLAe was more effective on resistant MRA-1240 than on the sensitive MRA-1252 strain. Because results were not as robust as observed in animal and human studies, a host interaction was suspected, so sera collected from adult and pediatric Kenyan malaria patients was used in RSA inhibition experiments and compared to sera from adults naïve to the disease. The sera from both age groups of malaria patients inhibited parasite growth ≥ 70% after treatment with DLAe and compared to malaria naïve subjects suggesting some host interaction with DLA. The discrepancy between these data and in-vivo reports suggested that DLA’s effects require an interaction with the host to unlock their potential as an antimalarial therapy. Although we showed there are serum-based host effects that can kill up to 95% of parasites in vitro, it remains unclear how or if they play a role in vivo. These results further our understanding of how DLAe works against the malaria parasite in vitro.

Mis en ligne par La vie re-belle
 29/03/2020
 https://lavierebelle.org/artemisia-annua-l-etudes-in-vitro

 Documents

 Antimalarial action of aqueous and hydro alcoholic extracts of {Artemisia annua} L . cultivated in Benin : In vitro and in vivo studies
PDF 
 Journal of Chemical and Pharmaceutical Research
 Etude préliminaire Sur l’effet des extraits de feuilles d’Artemisia annua et de Moringa oleifera sur Toxoplasmagondii in vitro
PDF 
 Ethnopharmacology in overdrive : The remarkable anti-HIV activity of Artemisia annua
PDF Journal of Ethnopharmacology. Elsevier
 Leishmanicidal activities of Artemisia annua leaf essential oil against Visceral Leishmaniasis
PDF 
 Frontiers in Microbiology

Etude in vitro d’Artemisia annua L.

Ce dossier regroupe les publications relatives aux activités d’Artemisia annua L. in vitro

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