Matthew R Desrosiers  1 Alexis Mittelman  2 Pamela J Weathers  1 Affiliations


Artemisia annua L. and artemisinin, have been used for millennia to treat malaria. We used human liver microsomes (HLM) and rats to compare hepatic metabolism, tissue distribution, and inflammation attenuation by dried leaves of A. annua (DLA) and pure artemisinin. For HLM assays, extracts, teas, and phytochemicals from DLA were tested and IC50 values for CYP2B6 and CYP3A4 were measured. For tissue distribution studies, artemisinin or DLA was orally delivered to rats, tissues harvested at 1 h, and blood, urine and feces over 8 h; all were analyzed for artemisinin and deoxyartemisinin by GC-MS. For inflammation, rats received an intraperitoneal injection of water or lipopolysaccharide (LPS) and 70 mg/kg oral artemisinin as pure drug or DLA. Serum was collected over 8 h and analyzed by ELISA for TNF-α, IL-6, and IL-10. DLA-delivered artemisinin distributed to tissues in higher concentrations in vivo, but elimination remained mostly unchanged. This seemed to be due to inhibition of first-pass metabolism by DLA phytochemicals, as demonstrated by HLM assays of DLA extracts, teas and phytochemicals. DLA was more effective than artemisinin in males at attenuating proinflammatory cytokine production; the data were less conclusive in females. These results suggest that the oral consumption of artemisinin as DLA enhances the bioavailability and anti-inflammatory potency of artemisinin.

Keywords: Artemisia annua; artemisinin; cytochrome P450 inhibition; inflammation; tissue distribution.

Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.


figure 1
Figure 1 Chemical structures of artemisinin (A) and deoxyartemisinin (B).
Figure 2 Artemisinin distribution in tissues and serum of male (A) and female (B) rats after oral delivery of artemisinin as dried leaf A. annua (DLA) or pure artemisinin. n = 3, *; p ≤ 0.05; error bars = SEM.
Figure 3 Deoxyartemisinin (deoxyAN) distribution in tissues and serum of male (A) and female (B) rats after oral delivery of artemisinin as dried leaf A. annua (DLA) or pure artemisinin. n = 3, *; p ≤ 0.05; error bars = SEM.
Figure 4 Artemisinin (AN) and deoxyartemisinin (DeoxyAN) accumulation in urine of male (A,B) and female (C,D) rats after oral delivery of artemisinin as dried leaf A. annua (DLA) or pure artemisinin. n = 8, *; p ≤ 0.05; error bars = SEM.
Figure 5 Artemisinin (A) and deoxyartemisinin (B) found in the urine of male and female rats orally gavaged with either pure artemisinin or dried leaves of A. annua (DLA). n = 8; * p ≤ 0.05; error bars = SEM.
fig 6
Figure 6 Production of proinflammatory cytokines TNF-α and IL-6 in male (A,C) and female (B,D) rats after LPS challenge and treatment with either pure artemisinin (AN) or DLA (equal artemisinin doses). n = 5–6 for experimental conditions, n = 3 for injection control; error bars = SEM; *, p ≤ 0.05 when comparing LPS+DLA to LPS Control; #, p ≤ 0.05 when comparing LPS+AN to LPS Control.


  1. WHO Malaria. [(accessed on 23 December 2019)]; Available online:
  2. WHO Guidelines for the treatment of malaria. [(accessed on 23 December 2019)]; Available online:
  3. Olsson M.E., Olofsson L.M., Lindahl A.L., Lundgren A., Brodelius M., Brodelius P.E. Localization of enzymes of artemisinin biosynthesis to the apical cells of glandular secretory trichomes of Artemisia annua L. Phytochemistry. 2009;70:1123–1128. doi: 10.1016/j.phytochem.2009.07.009. – DOIPubMed
  4. Hsu E. The history of qing hao in the Chinese materia medica. Trans. R. Soc. Trop. Med. Hyg. 2006;100:505–508. doi: 10.1016/j.trstmh.2005.09.020. – DOIPubMed
  5. Munyangi J., Cornet-Vernet L., Idumbo M., Lu C., Lutgen P., Perronne C., Ngombe N., Bianga J., Mupenda B., Lalukala P., et al. Artemisia annua and Artemisia afra tea infusions vs. artesunate-amodiaquine (ASAQ) in treating Plasmodium falciparum malaria in a large scale, double blind, randomized clinical trial. Phytomedicine. 2019;57:49–56. doi: 10.1016/j.phymed.2018.12.002. – DOIPMCPubMed
  6. ICIPE . Whole-leaf Artemisia Annua-based Antimalarial Drug: Report on Proof-of-Concepts Studies. ICIPE; Nairobi, Kenya: 2005.
  7. Desrosiers M.R., Weathers P.J. Effect of leaf digestion and artemisinin solubility for use in oral consumption of dried Artemisia annua leaves to treat malaria. J. Ethnopharmacol. 2016;190:313–318. doi: 10.1016/j.jep.2016.06.041. – DOIPMCPubMed
  8. Desrosiers M.R., Weathers P.J. Artemisinin permeability via Caco-2 cells increases after simulated digestion of Artemisia annua leaves. J. Ethnopharmacol. 2018;210:254–259. doi: 10.1016/j.jep.2017.08.038. – DOIPMCPubMed
  9. Weathers P.J., Arsenault P.R., Covello P.S., McMickle A., Teoh K.H., Reed D.W. Artemisinin production in Artemisia annua: Studies in planta and results of a novel delivery method for treating malaria and other neglected diseases. Phytochem. Rev. 2011;10:173–183. doi: 10.1007/s11101-010-9166-0. – DOIPMCPubMed
  10. Svensson U.S.H., Ashton M. Identification of the human cytochrome P450 enzymes involved in the in vitro metabolism of artemisinin. Br. J. Clin. Pharmacol. 1999;48:528–535. doi: 10.1046/j.1365-2125.1999.00044.x. – DOIPMCPubMed
  11. Lee I.S., Hufford C.D. Metabolism of antimalarial sesquiterpene lactones. Pharmacol. Ther. 1990;48:345–355. doi: 10.1016/0163-7258(90)90053-5. – DOIPubMed
  12. Weathers P.J., Elfawal M.A., Towler M.J., Acquaah-Mensah G.K., Rich S.M. Pharmacokinetics of artemisinin delivered by oral consumption of Artemisia annua dried leaves in healthy vs. Plasmodium chabaudi-infected mice. J. Ethnopharmacol. 2014;153:732–736. doi: 10.1016/j.jep.2014.03.037. – DOIPMCPubMed
  13. Räth K., Taxis K., Walz G., Gleiter C.H., Li S.M., Heide L. Pharmacokinetic study of artemisinin after oral intake of a traditional preparation of Artemisia annua L.(annual wormwood) Am. J. Trop. Med. Hyg. 2004;70:128–132. doi: 10.4269/ajtmh.2004.70.128. – DOIPubMed
  14. Xinyi N., Liyi H.O., Zhihong R., Zhenyu S. Metabolic fate of Qinghaosu in rats; a new TLC densitometric method for its determination in biological material. Eur. J. Drug Metab. Pharmacokinet. 1985;10:55–59. doi: 10.1007/BF03189697. – DOIPubMed
  15. Ashton M. Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat. Xenobiotica. 1999;29:195–204. doi: 10.1080/004982599238740. – DOIPubMed
  16. Wang K.S., Li J., Wang Z., Mi C., Ma J., Piao L.X., Xu G.H., Li X., Jin X. Artemisinin inhibits inflammatory response via regulating NF-κB and MAPK signaling pathways. Immunopharmacol. Immunotoxicol. 2017;39:28–36. doi: 10.1080/08923973.2016.1267744. – DOIPubMed
  17. Kim W.S., Choi W.J., Lee S., Kim W.J., Lee D.C., Sohn U.D., Shin H.S., Kim W. Anti-inflammatory, Antioxidant and Antimicrobial Effects of Artemisinin Extracts from Artemisia annua L. Kor. J. Physiol. Pharmacol. 2015;19:21–27. doi: 10.4196/kjpp.2015.19.1.21. – DOIPMCPubMed
  18. Rathee P., Chaudhary H., Rathee S., Rathee D., Kumar V., Kohli K. Mechanism of action of flavonoids as anti-inflammatory agents: a review. Inflamm. Allergy Drug Targets. 2009;8:229–235. doi: 10.2174/187152809788681029. – DOIPubMed
  19. De Cássia da Silveira e Sá R., Andrade L.N., de Sousa D.P. A review on anti-inflammatory activity of monoterpenes. Molecules. 2013;18:1227–1254. doi: 10.3390/molecules18011227. – DOIPMCPubMed
  20. Li Y.J., Guo Y., Yang Q., Weng X.G., Yang L., Wang Y.J., Chen Y., Zhang D., Li Q., Liu X.C., et al. Flavonoids casticin and chrysosplenol D from Artemisia annua L. inhibit inflammation in vitro and in vivo. Toxicol. Appl. Pharmacol. 2015;286:151–158. doi: 10.1016/j.taap.2015.04.005. – DOIPubMed
  21. Juergens U.R., Engelen T., Racké K., Stöber M., Gillissen A., Vetter H. Inhibitory activity of 1, 8-cineol (eucalyptol) on cytokine production in cultured human lymphocytes and monocytes. Pulm. Pharmacol. Ther. 2004;17:281–287. doi: 10.1016/j.pupt.2004.06.002. – DOIPubMed
  22. Melillo de Magalhães P., Dupont I., Hendrickx A., Joly A., Raas T., Dessy S., Sergent T., Schneider Y.-J. Anti-inflammatory effect and modulation of cytochrome P450 activities by Artemisia annua tea infusions in human intestinal Caco-2 cells. Food Chem. 2012;134:864–871. doi: 10.1016/j.foodchem.2012.02.195. – DOIPubMed
  23. Weathers P.J., Jordan N.J., Lasin P., Towler M.J. Simulated digestion of dried leaves of Artemisia annua consumed as a treatment (pACT) for malaria. J. Ethnopharmacol. 2014;151:858–863. doi: 10.1016/j.jep.2013.11.043. – DOIPMCPubMed
  24. Weathers P.J., Towler M.J. The flavonoids casticin and artemetin are poorly extracted and are unstable in an Artemisia annua tea infusion. Planta Med. 2012;78:1024–1026. doi: 10.1055/s-0032-1314949. – DOIPMCPubMed
  25. Arvouet-Grand A., Vennat B., Pourrat A., Legret P. Standardization of propolis extract and identification of principal constituents. J. Pharm. Belg. 1994;49:462–468. – PubMed
  26. Weathers P.J., Towler M.J. Changes in key constituents of clonally propagated Artemisia annua L. during preparation of compressed leaf tablets for possible therapeutic use. Ind. Crop. Prod. 2014;62:173–178. doi: 10.1016/j.indcrop.2014.08.033. – DOIPMCPubMed
  27. Gruessner B.M., Cornet-Vernet L., Desrosiers M.R., Lutgen P., Towler M.J., Weathers P.J. It is not just artemisinin: Artemisia sp. for treating diseases including malaria and schistosomiasis. Phytochem. Rev. 2019 doi: 10.1007/s11101-019-09645-9. – DOI
  28. Munyangi J., Cornet-Vernet L., Idumbo M., Lu C., Lutgen P., Perronne C., Ngombe N., Bianga J., Mupenda B., Lalukala P., et al. Effect of Artemisia annua and Artemisia afra tea infusions on schistosomiasis in a large clinical trial. Phytomedicine. 2018;51:233–240. doi: 10.1016/j.phymed.2018.10.014. – DOIPMCPubMed
  29. Titulaer H.A.C., Zuidema J., Kager P.A., Wetsteyn J., Lugt C.B., Merkus F. The pharmacokinetics of artemisinin after oral, intramuscular and rectal administration to volunteers. J. Pharm. Pharmacol. 1990;42:810–813. doi: 10.1111/j.2042-7158.1990.tb07030.x. – DOIPubMed
  30. Towler M.J., Weathers P.J. Variations in key artemisinic and other metabolites throughout plant development in Artemisia annua L. for potential therapeutic use. Ind. Crop. Prod. 2015;67:185–191. doi: 10.1016/j.indcrop.2015.01.007. – DOIPMCPubMed
  31. Bison S., Carboni L., Arban R., Bate S., Gerrard P.A., Razzoli M. Differential behavioral, physiological, and hormonal sensitivity to LPS challenge in rats. Int. J. Interferon Cytokine Mediat. Res. 2009;1:1–13. doi: 10.2147/IJICMR.S4273. – DOI
  32. Ferreira J.F.S., Luthria D.L., Sasaki T., Heyerick A. Flavonoids from Artemisia annua L. as antioxidants and their potential synergism with artemisinin against malaria and cancer. Molecules. 2010;15:3135–3170. doi: 10.3390/molecules15053135. – DOIPMCPubMed
  33. Lau A.J., Chang T.K.H. Inhibition of Human CYP2B6-Catalyzed Bupropion Hydroxylation by Ginkgo biloba Extract: Effect of Terpene Trilactones and Flavonols. Drug Metab. Dispos. 2009;37:1931. doi: 10.1124/dmd.109.028118. – DOIPubMed
  34. Wei S., Ji H., Yang B., Ma L., Bei Z., Li X., Dang H., Yang X., Liu C., Wu X., et al. Impact of chrysosplenetin on the pharmacokinetics and anti-malarial efficacy of artemisinin against Plasmodium berghei as well as in vitro CYP450 enzymatic activities in rat liver microsome. Malar. J. 2015;14:432. doi: 10.1186/s12936-015-0929-3. – DOIPMCPubMed
  35. Sergent T., Dupont I., Van Der Heiden E., Scippo M.L., Pussemier L., Larondelle Y., Schneider Y.J. CYP1A1 and CYP3A4 modulation by dietary flavonoids in human intestinal Caco-2 cells. Toxicol. Lett. 2009;191:216–222. doi: 10.1016/j.toxlet.2009.09.002. – DOIPubMed
  36. Choi J.S., Piao Y.J., Kang K.W. Effects of quercetin on the bioavailability of doxorubicin in rats: Role of CYP3A4 and P-gp inhibition by quercetin. Arch. Pharm. Res. 2011;34:607–613. doi: 10.1007/s12272-011-0411-x. – DOIPubMed
  37. Alin M.H., Bjorkman A. Concentration and time dependency of artemisinin efficacy against Plasmodium falciparum in vitro. Am. J. Trop. Med. Hyg. 1994;50:771–776. doi: 10.4269/ajtmh.1994.50.771. – DOIPubMed
  38. WHO Position Statement. Effectiveness of Non-Pharmaceutical Forms of Artemisia annua L. against malaria. [(accessed on 23 December 2019)]; Available online:
  39. WHO The Use of Non-pharmaceutical Forms of Artemisia. [(accessed on 23 December 2019)]; Available online: