In vitro and in vivo antileishmanial activity of Artemisia annua L. leaf powder and its potential usefulness in the treatment of uncomplicated cutaneous leishmaniasis in humans

ABSTRACT INTRODUCTION: Cutaneous leishmaniasis (CL) is a tropical disease that affects millions of individuals worldwide. The current drugs for CL may be effective but have serious side effects; hence, alternatives are urgently needed. Although plant-derived materials are used for the treatment of various diseases in 80% of the global population, the validation of these products is essential. Gelatin capsules containing dried Artemisia annua leaf powder were recently developed as a new herbal formulation (totum) for the oral treatment of malaria and other parasitic diseases. Here, we aimed to determine the usefulness of A. annua gel capsules in CL. METHODS: The antileishmanial activity and cytotoxicity of A. annua L. capsules was determined via in vitro and in vivo studies. Moreover, a preliminary evaluation of its therapeutic potential as antileishmanial treatment in humans was conducted in 2 patients with uncomplicated CL. RESULTS: Artemisia annua capsules showed moderate in vitro activity in amastigotes of Leishmania (Viannia) panamensis; no cytotoxicity in U-937 macrophages or genotoxicity in human lymphocytes was observed. Five of 6 (83.3%) hamsters treated with A. annua capsules (500mg/kg/day) for 30 days were cured, and the 2 examined patients were cured 45 days after initiation of treatment with 30g of A. annua capsules, without any adverse reactions. Both patients remained disease-free 26 and 24 months after treatment completion. CONCLUSION: Capsules of A. annua L. represent an effective treatment for uncomplicated CL, although further randomized controlled trials are needed to validate its efficacy and safety.

Atividade ovicida e larvicida do extrato hidroalcoólico de Artemisia annua sobre parasitas gastrintestinais de bovinos / Ovicidal and larvicidal activity of Artemisia annua hydroalcoholic extract against gastrointestinal parasites in cattle

Os parasitas gastrintestinais causam enorme prejuízo econômico na bovinocultura, tanto nacional como mundial, ocasionado principalmente por Bunostumom sp., Cooperia sp. e Trichostrongylus sp. O objetivo deste trabalho foi determinar a eficácia in vitro do extrato hidroalcoólico de Artemisia annua (H.7) frente a esses endoparasitas. O H.7 foi produzido com sete dias de percolação a 4ºC e posteriormente liofilizado. Com esse fitoterápico, realizaram-se testes de eclodibilidade de ovos (TEO) e de migração larvar em ágar (TMLA), com seis repetições, com concentrações crescentes (0,78 a 50mg/mL). Para analisar a composição química do fitoterápico, procedeu-se à marcha fitoquímica completa. No TEO, a eficácia variou de 94,08±2,58% na maior concentração a 15,67±0,97% na menor concentração. Já no TMLA os valores encontrados variaram de 90,05±0,55% a 4,12±0,46%. Nas análises fitoquímicas, foram encontrados diversos compostos com propriedades de combater os nematódeos, tanto direta como indiretamente. Os resultados obtidos nos testes in vitro evidenciam que o extrato produzido possui potencial de combater nematódeos gastrintestinais de bovinos. Novos estudos devem ser realizados buscando maximizar a eficácia do H.7 e de outras extrações obtidas a partir de A. annua, uma vez que foram demonstrados excelentes resultados em ambos os experimentos.

Gastrointestinal parasites cause economic losses to the cattle production, in Brazil and worldwide, mainly caused by Bunostumom sp., Cooperia sp. and Trichostrongylus sp. The aim of this study was to determine the in vitro efficacy of hydroalcoholic extract of Artemisia annua (H.7) against these parasites. The H.7 was produced after 7 days of storage at 4°C and then lyophilized. With this herbal the egg hatch test (EHT) and larval migration inhibition (LMI) were performed,in six replicates with different concentrations (0.78 to 50mg/mL). To analyze the chemistry composition the complete phytochemical screening was done. In EHT efficiency ranged from 94.08±2.58% at the highest concentration to 15.67± 0.97% in the lowest concentration. In LMI test the values ranged from 90.05±0.55% to 4.12±0.46%. Phytochemical tests showed many chemical compounds with anthelmintic properties. The results obtained in biochemical tests together with those found in in vitro tests showed that the extract produced has the potential to combat intestinal nematodes of cattle. Further studies should be conducted to maximize the effectiveness of H.7 and other extractions from A. annua, because it demonstrated excellent results in both experiments.

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

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 P. falciparum K1 and 3d7 strains and field isolates from the state of Amazonas, Brazil, to A. annua infusions (5 g dry leaves in 1 L of boiling water) and the drug standards chloroquine, quinine and artemisinin were evaluated. The A. 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 A. annua leaves used were 0.90-1.13% (m/m). The concentration of artemisinin in the infusions was 40-46 mg/L. Field P. falciparum isolates were resistant to chloroquine and sensitive to quinine and artemisinin. The average 50% inhibition concentration values for A. 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 P. falciparum due to their artemisinin concentration. No synergistic effect was observed for artemisinin in the infusions.

Effective method for extraction of larvicidal component from leaves of Azadirachta indica and Artemisia annua Linn.

Leaves of Artemisia annua linn. and Azadirachta indica were extracted in petroleum ether and hexane respectively by different methods of extraction i.e. cold extraction, reflux extraction and soxhlet extraction. The crude extract obtained was tested against third instar larvae of Anopheles stephensi. On comparison of larval mortality of crude extract obtained by these three methods, both soxhlet and reflux extraction method showed 100% mortality at 200 ppm after 48 hr in case of A. annua. However LC50 (20 ppm) value of crude extract obtained by soxhlet extraction showed better results than reflux extraction (35 ppm) method after 72 hr. In case of A. indica, crude obtained by soxhlet showed 100% mortality (after 48 hr) at 250 ppm and LC50 of 69 ppm at 72 hr. Reflux extraction does not show any appreciable mortality even at 250 ppm concentration.