Cited by 366Plant biochemistry and biosynthesisSesquiterpenes and Asteraceae StudiesPhytochemical compounds biological activities

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The Biosynthesis of Artemisinin (Qinghaosu) and the Phytochemistry of Artemisia annua L. (Qinghao)

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Brown, Geoffrey D.·2010·Molecules

Abstract

The Chinese medicinal plant Artemisia annua L. (Qinghao) is the only known source of the sesquiterpene artemisinin (Qinghaosu), which is used in the treatment of malaria. Artemisinin is a highly oxygenated sesquiterpene, containing a unique 1,2,4-trioxane ring structure, which is responsible for the antimalarial activity of this natural product. The phytochemistry of A. annua is dominated by both sesquiterpenoids and flavonoids, as is the case for many other plants in the Asteraceae family. However, A. annua is distinguished from the other members of the family both by the very large number of natural products which have been characterised to date (almost six hundred in total, including around fifty amorphane and cadinane sesquiterpenes), and by the highly oxygenated nature of many of the terpenoidal secondary metabolites. In addition, this species also contains an unusually large number of terpene allylic hydroperoxides and endoperoxides. This observation forms the basis of a proposal that the biogenesis of many of the highly oxygenated terpene metabolites from A. annua – including artemisinin itself – may proceed by spontaneous oxidation reactions of terpene precursors, which involve these highly reactive allyllic hydroperoxides as intermediates. Although several studies of the biosynthesis of artemisinin have been reported in the literature from the 1980s and early 1990s, the collective results from these studies were rather confusing because they implied that an unfeasibly large number of different sesquiterpenes could all function as direct precursors to artemisinin (and some of the experiments also appeared to contradict one another). As a result, the complete biosynthetic pathway to artemisinin could not be stated conclusively at the time. Fortunately, studies which have been published in the last decade are now providing a clearer picture of the biosynthetic pathways in A. annua. By synthesising some of the sesquiterpene natural products which have been proposed as biogenetic precursors to artemisinin in such a way that they incorporate a stable isotopic label, and then feeding these precursors to intact A. annua plants, it has now been possible to demonstrate that dihydroartemisinic acid is a late-stage precursor to artemisinin and that the closely related secondary metabolite, artemisinic acid, is not (this approach differs from all the previous studies, which used radio-isotopically labelled precursors that were fed to a plant homogenate or a cell-free preparation). Quite remarkably, feeding experiments with labeled dihydroartemisinic acid and artemisinic acid have resulted in incorporation of label into roughly half of all the amorphane and cadinane sesquiterpenes which were already known from phytochemical studies of A. annua. These findings strongly support the hypothesis that many of the highly oxygenated sesquiterpenoids from this species arise by oxidation reactions involving allylic hydroperoxides, which seem to be such a defining feature of the chemistry of A. annua. In the particular case of artemisinin, these in vivo results are also supported by in vitro studies, demonstrating explicitly that the biosynthesis of artemisinin proceeds via the tertiary allylic hydroperoxide, which is derived from oxidation of dihydroartemisinic acid. There is some evidence that the autoxidation of dihydroartemisinic acid to this tertiary allylic hydroperoxide is a non-enzymatic process within the plant, requiring only the presence of light; and, furthermore, that the series of spontaneous rearrangement reactions which then convert this allylic hydroperoxide to the 1,2,4-trioxane ring of artemisinin are also non-enzymatic in nature.

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TL;DR

This review compiles the phytochemistry of *Artemisia annua* and argues that many highly oxygenated terpenes, including artemisinin, arise via spontaneous autoxidation of terpene precursors through allylic hydroperoxides. It concludes that dihydroartemisinic acid, not artemisinic acid, is the late-stage precursor to artemisinin, with the final steps likely non-enzymatic.

Key findings

  • Nearly 600 secondary metabolites have been characterized from *A. annua*, including around 50 amorphane and cadinane sesquiterpenes; artemisinin content varies from 0.01% to 1%, and can reach 1.4% in some cultivated strains.
  • Stable-isotope feeding to intact plants showed dihydroartemisinic acid is a late-stage precursor to artemisinin, whereas artemisinic acid is not; labeled dihydroartemisinic acid yielded sixteen labeled amorphane/cadinane sesquiterpenes, while labeled artemisinic acid yielded seven metabolites.
  • Artemisinin biosynthesis is proposed to proceed via the tertiary allylic hydroperoxide derived from dihydroartemisinic acid; in vitro evidence supports a four-step spontaneous autoxidation mechanism, and autoxidation to the hydroperoxide may require only light and be non-enzymatic.
  • Many highly oxygenated terpenes from *A. annua* are proposed to arise by oxidation reactions involving allylic hydroperoxides and endoperoxides, which the review treats as a defining feature of this species’ chemistry.

Methods

  • Comprehensive review of original phytochemical literature on *A. annua* up to 2009, organizing almost 600 secondary metabolites into eight sections and more than forty tables using a largely biogenetic classification.
  • Synthesizes biosynthetic studies including older radiolabeled cell-free/homogenate experiments and more recent stable-isotope feeding to intact plants, alongside in vitro autoxidation experiments.
  • Evaluates three phases of artemisinin biosynthesis and production strategies: plant breeding, plant tissue culture, endophytic fungi, and genetic engineering.

Limitations

  • The paper states that the proposed biosynthetic route is still not universally accepted and that older literature contains results that may appear to contradict it; many phase 3 intermediates have been proposed, and not all can be correct.
  • No enzymes have yet been described for phase 3 reactions, and there is no direct in vivo evidence for steps ii–iv of the proposed autoxidation mechanism; evidence for those steps comes solely from in vitro studies.
  • The text notes that understanding of spontaneous autoxidation without a photosensitizer remains incomplete; as a review, conclusions depend on synthesis of prior literature rather than a single new experimental dataset (evident from the text).

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Crossref · 10.3390/molecules15117603

Cites

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Figures

Figure 1 Artemisinin and its semi-synthetic derivatives, which are currently used in the treatment of malaria.
Figure 1 Artemisinin and its semi-synthetic derivatives, which are currently used in the treatment of malaria.CC BY · PubMed Central
Figure 2 Structures of isopentenyl pyrophosphate (IPP), dimethylallyl pyrophosphate (DMAPP), geranyl pyrophosphate (GPP), neryl pyrophosphate (NPP) and linaloyl pyrophosphate (LPP) which are all possible precursors to monoterpenes from A.annua.
Figure 2 Structures of isopentenyl pyrophosphate (IPP), dimethylallyl pyrophosphate (DMAPP), geranyl pyrophosphate (GPP), neryl pyrophosphate (NPP) and linaloyl pyrophosphate (LPP) which are all possible precursors to monoterpenes from A.annua.CC BY · PubMed Central
Scheme 1 Postulated biosynthesis of allylic hydroperoxides: α-myrcene hydroperoxide (265) and β-myrcene hydroperoxide (264) via spontaneous autoxidation reactions at the tri-substituted bond of the precursor myrcene (254). Further reactions of such allylic hydroperoxides would account for the biogenesis of oxygenated monoterpenes such as (267), (263), (271) and (272).
Scheme 1 Postulated biosynthesis of allylic hydroperoxides: α-myrcene hydroperoxide (265) and β-myrcene hydroperoxide (264) via spontaneous autoxidation reactions at the tri-substituted bond of the precursor myrcene (254). Further reactions of such allylic hydroperoxides would account for the biogenesis of oxygenated monoterpenes such as (267), (263), (271) and (272).CC BY · PubMed Central
Scheme 2 Formation of the irregular artemisyl, lavandulyl and santolinyl skeletons in A. annua by “head-to-middle” condensation of a DMAPP (C5) precursor and subsequent carbon-carbon cleavage reactions of the resulting intermediate, chryanthemyl pyrophosphate.
Scheme 2 Formation of the irregular artemisyl, lavandulyl and santolinyl skeletons in A. annua by “head-to-middle” condensation of a DMAPP (C5) precursor and subsequent carbon-carbon cleavage reactions of the resulting intermediate, chryanthemyl pyrophosphate.CC BY · PubMed Central
Scheme 3 Proposed formation of lavandulanes 279 and 280 by spontaneous autoxidation reactions.
Scheme 3 Proposed formation of lavandulanes 279 and 280 by spontaneous autoxidation reactions.CC BY · PubMed Central
Scheme 4 Postulated biogenesis of (414) and (413) by spontaneous autoxidation reaction of (422), followed by reduction.
Scheme 4 Postulated biogenesis of (414) and (413) by spontaneous autoxidation reaction of (422), followed by reduction.CC BY · PubMed Central
Scheme 5 Proposed biogenesis of arteannuin H (465) and other amorphane sesquiterpenes from A. annua via tertiary and allylic secondary hydroperoxides which are derived from spontaneous autoxidation of dihydroartemisinic acid (480)/artemisinic acid (473).
Scheme 5 Proposed biogenesis of arteannuin H (465) and other amorphane sesquiterpenes from A. annua via tertiary and allylic secondary hydroperoxides which are derived from spontaneous autoxidation of dihydroartemisinic acid (480)/artemisinic acid (473).CC BY · PubMed Central
Scheme 6 The most dominant products from metabolism of dihydroartemisinic acid (480) in vivo in A. annua plants.
Scheme 6 The most dominant products from metabolism of dihydroartemisinic acid (480) in vivo in A. annua plants.CC BY · PubMed Central
Scheme 7 The most dominant products from metbolism of artemisinic acid (473) in vivo in A. annua plants.
Scheme 7 The most dominant products from metbolism of artemisinic acid (473) in vivo in A. annua plants.CC BY · PubMed Central
Scheme 8 Proposed formation of phytene-1-ol-2-hydroperoxide (553) by spontaneous autoxidation of phytol (550) and subsequent homolysis/reduction of 553 to phytene-1,2-diol (552).
Scheme 8 Proposed formation of phytene-1-ol-2-hydroperoxide (553) by spontaneous autoxidation of phytol (550) and subsequent homolysis/reduction of 553 to phytene-1,2-diol (552).CC BY · PubMed Central
Scheme 9 Three phases in the biosynthesis of artemisinin (495).
Scheme 9 Three phases in the biosynthesis of artemisinin (495).CC BY · PubMed Central
Scheme 10 Cyclization of FPP (378) to amorpha-4,11-diene (451), catalysed by the enzyme ADS.
Scheme 10 Cyclization of FPP (378) to amorpha-4,11-diene (451), catalysed by the enzyme ADS.CC BY · PubMed Central
Figure 3 Minor products from the cyclization of FPP (378) by ADS, which have not yet been reported as natural products from A. annua. Amorph-4-en-11-ol (574) is probably formed by quenching of the C-11 amorphane cation by water; β-sesquiphellandrene (575) by elimination of H-15 from the C-1 bisabolyl cation; zingiberene (576) from elimination of H-4 from the C-1 biasabolyl cation; zingiberenol (577) by quenching of the allylic C-1 bisabolyl cation by water; and γ-humulene (578) by an alternative cyclization of nerolidyl pyrophosphate.
Figure 3 Minor products from the cyclization of FPP (378) by ADS, which have not yet been reported as natural products from A. annua. Amorph-4-en-11-ol (574) is probably formed by quenching of the C-11 amorphane cation by water; β-sesquiphellandrene (575) by elimination of H-15 from the C-1 bisabolyl cation; zingiberene (576) from elimination of H-4 from the C-1 biasabolyl cation; zingiberenol (577) by quenching of the allylic C-1 bisabolyl cation by water; and γ-humulene (578) by an alternative cyclization of nerolidyl pyrophosphate.CC BY · PubMed Central
Scheme 11 Various possible routes for the oxidation of the isopropylidene group in amorpha-4,11-diene (451), yielding artemisinic acid (473) and/or dihydroartemisinic acid (480) in phase 2 of the biosynthesis of artemisinin.
Scheme 11 Various possible routes for the oxidation of the isopropylidene group in amorpha-4,11-diene (451), yielding artemisinic acid (473) and/or dihydroartemisinic acid (480) in phase 2 of the biosynthesis of artemisinin.CC BY · PubMed Central
Scheme 12 A four-step mechanism for the spontaneous autoxidation of dihydroartemisinic acid (480) to artemisinin (495) in A. annua.
Scheme 12 A four-step mechanism for the spontaneous autoxidation of dihydroartemisinic acid (480) to artemisinin (495) in A. annua.CC BY · PubMed Central
Figure 4 Metabolites which have been isolated from tissue culture of A.annua, but which have not yet been reported from A. annua plants.
Figure 4 Metabolites which have been isolated from tissue culture of A.annua, but which have not yet been reported from A. annua plants.CC BY · PubMed Central
Scheme 13 Genetic engineering of enzymes for the production of artemisinic acid (473) from A. annua in to a microbial host.
Scheme 13 Genetic engineering of enzymes for the production of artemisinic acid (473) from A. annua in to a microbial host.CC BY · PubMed Central

Verification

Claims the paper makes

169 Not checked yet · 51 No basis given · 42 Own interpretation · 11 Own result

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Show all 136 claims

  • Probably not a substantial claim. 104
  • It may state two separate claims as one. 55
137 of 137
  • The authors state this as an assumption.

    The final steps in the biosynthetic pathway to artemisinin proceed from dihydroartemisinic acid (480), rather than artemisinic acid (473) or other proposed late-stage intermediates.

    “In this review, it is assumed that the final steps in the biosynthetic pathway to artemisinin (495) proceed from dihydroartemisinic acid (480), rather than artemisinic acid (473) (or, indeed, any of the other late stage intermediates: 462, 478, 479, 485, 494 or 497 which have been proposed in the past).”
  • The authors state this as an assumption.

    The configuration at the 7- and 11-positions of phytene-1,2-diol (552) remained fixed, as in phytol.

    “on the assumption that the configuration at the 7- and 11- positions of (552) remained fixed, as in phytol.”
  • The authors state this as an assumption.

    Artemisinic acid (473) is a “dead end” metabolite, leading away from artemisinin (495) in phase 2 of the biosynthetic route.

    “the existence of these two chemotypes might instead be explained on the assumption that artemisinic acid (473) is a “dead end” metabolite, leading “away” from artemisinin (495) in phase 2 of the biosynthetic route, as is implied in Scheme 11.”
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Citations

Cites

Show all 328 references

Research works printed but not linked: 81

  1. (1992). Pharmacopoeia of the People’s Republic of China, English edition ed., Guangdong Science and Technology Press.
  2. Balachandran, 1987, Chemical investigation of some Artemisia species: Search for artemisinin or other related sesquiterpene lactones with a peroxide bridge, Indian J. Pharm. Sci., 49, 152
  3. Bhakuni, 2001, Secondary metabolites of Artemisia annua and their biological activity, Curr. Sci., 80, 35
  4. Buckingham, 2004, Introduction to the Type of Compound Index, Dictionary of Natural Products, Volume 1, xi
  5. Tian, 2007, Study on volatile constituents of herba Artemisia annua and its preparation by GC-MS, Shizen Guoyi Guoyao, 18, 1840
  6. Zou, 1999, Analysis of volatile components from Artemisia annua Linn, Fenxi Ceshi Xuebao, 18, 55
  7. Zhong, 1983, Studies of volatile constituents from Artemisia annua, Zhongyao Tongbao, 8, 31
  8. Genov, 1983, Gas-Chromatographic study of the essential oil of Artemisia annua L., Naunchi Trudove - Vissh Institut po Khranitelna i Vkusova Promishlenost, Plodiv, 30, 141
  9. Tsankova, 1976, On the composition of the essential oil from Artemisia annua Linnaeus, Italiana Essenze, Profumi, Piante Officinale, Aromi, Saponi, Cosmetici, Aerosol, 58, 502
  10. Biougne, 1993, Essential oil of Artemisia annua: Seasonal variations in chemical composition, Rivista Italiania EPPOS, 4, 622
  11. Popescu, 1980, Artemisia annua: An indigenous source of volatile oil, Clujul Med., 53, 331
  12. He, 2008, Analysis of chemical composition of voatile oil in Youyang Artemisia annua by GC-MS, Huaxi Yaoxue Zazhi, 23, 30
  13. Zhang, 2004, Studies on the chemical composition of the essential oil of Artemisia annua L. from Xinjiang, Xibei Shifan Daxue Xuebao, Ziran Kexueban, 40, 67
  14. Tian, 1982, Studies on the chemical constituents of Qinghao (Artemisia annua), a traditional chinese herb, Zhongcaoyao, 13, 9
  15. Zhang, 2004, Studies on the composition of the fatty acids of five species of Artemisia in Xinjiang, Xibei Shifan Daxue Xuebao, Ziran Kexueban, 40, 64
  16. Deng, 2008, Alleochemicals isolation and structure identification of Artemisia annua, Huanan Nongye Daxue Xuebao, 29, 42
  17. Chen, 2001, Isolation and identification of arteannuin and its precursor, Zhongcaoyao, 32, 302
  18. Georgiev, 1983, Changes in the essential oil of Artemisia annua L. during storage, Nauchni Trudove - Vissh Institut po Khranitelna i Vkusova Promishlenost, 30, 149
  19. Georgiev, 1978, On the distillation of annual wormwood (Artemisia annua Linnaeus), Rivista Italiana Essenze, Profumi, Piante Officinali, Arromatizanti, Syndets, Saponi, Cosmaetici, Aerosols, 60, 302
  20. Tu, 1985, Chemical constituents of sweet wormwood (Artemisia annua) (III), Zhongcaoyao, 16, 200
  21. Liu, 1980, Studies on the constituents of Artemisia annua, Yaoxue Tongbao, 15, 39
  22. Djermanovic, 1997, Some new constituents from domestic plant species of Artemisia monogyna W and K and Artemisia annua L., J. Serb. Chem. Soc., 62, 113
  23. Yang, 2006, Determination of scopoletin in Artemisia annua by HPLC, Zhongguo Shiyan Fangjixue Zazhi, 12, 10
  24. Nguyen, 1998, Research on the extraction of a coumarin derivative from the leaves of Artemisia annua L. growing in Vietnam, Tap Chi Duoc Hoc, 36, 58
  25. Yang, 1989, Methoxylated flavones and coumarins from Artemisia annua, Phytochemistry, 28, 1509, 10.1016/s0031-9422(00)97776-8 · DOI
  26. Jeremic, 1979, Flavonols from Artemisia annua, Glas. Khem. Drush. Beograd., 44, 615
  27. Bhardwaj, 1985, Constituents of Artemisia annua, Proc. Indian Nat. Sci. Acad., 51A, 741
  28. Liu, 1981, Studies on the chemical constituents of Artemisia annua L., Yaoxue Xuebao (Acta Pharmaceutica Sinica), 16, 65
  29. Georgiev, 1981, Changes in yield and quality of essential oil in annual wormwood (Artemisia annua L.) during growth, Rastenievundi Nauki, 18, 95
  30. Hethelyl, 1994, Capillary gas chromatographic investigation of Artemisia annua essential oils, Olaj, Szappan, Kozmetika, 43, 103
  31. Youann, 1955, Essential oil of Artemisia annua, Perfumery and Essential Oil Rec., 46, 75
  32. Tu, 1981, Studies on the constituents of Artemisia annua, Yaoxue Xuebao (Acta Pharmaceutica Sinica), 16, 366
  33. Liu, 2007, Determination of camphor and caryophyllene in Artemisia annua L. and Herba artemisiae annuae oil by capillary GC, Huaxue Yaoxue Zashi, 22, 218
  34. Zhu, 1982, Study on antibacterial constituents of Qing Hao (Artemisia annua L.), Zhongcaoyao, 13, 6
  35. Zhou, 1987, Synthesis of arteannuin and related compounds, Yiyao Gongye, 18, 470
  36. Agrawal, 1996, NMR spectral investigations. Part 41. Sesquiterpenoids from Artemisia annua: 13C NMR shielding behavior, J. Sci. Ind. Res., 55, 17
  37. Tu, 1981, Chemical constituents in Artemisia annua L. and the derivatives of artemisinin, Chung Yao Tong Pao, 6, 31
  38. Pang, 1997, Study on the absolute configuration of artemisinic acid isolated from chinese antimalaria herb Artemisia annua L., Shenyang Yaoke Daxue Xuebao, 14, 189
  39. Zhou, 1989, Studies on the structure and synthesis of arteannuin and related compounds, Huaxue Xuebao, 47, 710
  40. Xu, 1982, Stereoselective synthesis of arteannuin B, Kexue Tongbao, 27, 1022
  41. Zhou, 1986, Studies on the structure and syntheses of arteannuin and related compounds XVIII. Synthesis of arteannuin A, Huaxue Xuebao, 44, 968
  42. Stefanovic, 1977, Chemical transformations of arteannuin B - cadinane sesquiterpenic lactone - isolated from Artemisia annua L., Glas. Khem. Drus., Beograd, 42, 227
  43. Pajarin, 1987, The total synthesis of (+/-)-arteannuin B, Diss. Abstr. Int., 48, 444
  44. Barriault, L., and Deon, D.H. (2001, January April). Recent progress towards the total synthesis of arteannuin M. Abstracts of Papers; In Proceedings of 221st ACS national Meeting, San Diego, CA, USA.
  45. Engel, D.A., Lam, H.T.C., Singletary, J.A., Poon, K.W.C., and Dudley, G.B. (2006, January March). Synthetic approaches to artemisinin-related natural products, dihydroartemisinic acid and arteannuin M. Abstracts of Papers; In Proceedings of 231st ACS national Meeting, Atlanta, GA, USA.
  46. Engel, D.A., Lam, H.T.C., Singletary, J.A., Poon, K.-W.C., and Dudley, G.B. (2008, January April). Synthetic approaches to artemisinin-related natural products, didesmethylartemisinin and dihydro-epi-deoxyarteannuin B. Abstracts of Papers; In Proceedings of 235th ACS national Meeting, New Orleans, LA, USA.
  47. Wu, 1984, Structure and synthesis of arteannuin and related compounds. XI, Identification of epoxyarteannuic acid, Huaxue Xuebao, 42, 596
  48. Kasymov, 1986, Lactones of Artemisia annua, Khim. Prir. Soedin., 5, 636
  49. Deng, 1981, Studies on the structure of the artemisic acid, Kexue Tongbao, 26, 1209
  50. Zhu, 1982, Studies on the antibacterial artemisinin, Zhongcaoyao (Chinese Traditional and Herbal Drugs), 13, 6
  51. Huang, 1987, Constituents of Artemisia annua L. I. Isolation and identification of 11R-(-)-dihydroarteannuic acid, Huaxue Xuebao, 45, 609
  52. Anonymous (1977). Co-ordinating group of research on the structure of Qing Hao Su. A new type of lactone - Qing Hau Sau. K’O Hsueh Tung Pao, 22, 142. (CA 1977, 87, 98788).
  53. Huang, 1988, Two-dimensional NMR studies of arteannuin, Huaxue Xuebao, 45, 305
  54. Leban, 1988, Crystal and molecular structure of Qinghaosu: A redetermination, Acta Pharmaceutica Jugoslavica, 38, 71
  55. Madhusudanan, 1989, Mass spectral studies on artemisinin, dihydroartemisinin and artether, Indian J. Chem. Sect. B, Organic Chemistry including Medicinal Chemistry, 28, 751
  56. Mi, 1987, Circular dichroism of some synthetic intermeidates of qinghaosu, Yaowu Fenxi Zazhi, 7, 262
  57. Wang, 1986, The 13C NMR spectroscopy of arteannuin analogues, Huaxue Xuebao, 44, 834
  58. Zhang, 1988, Structure and synthesis of arteannuin and related compounds. XXIV. Conversion of arteannuin into artemisitene, Youji Huaxue, 8, 329
  59. Gu, 1987, Pharmacological studies on artemisinin and its derivatives as antimalarial, Int. Congr. Ser. - Excerpta Med., 750, 657
  60. Shukla, 1991, Growth inhibitors from Artemisia annua, Indian Drugs, 28, 376
  61. Feng, 2004, Cloning and sequencing of Artemisia annua squalene synthase gene and its cDNA, Guangzhou Zhongyiyao Daxue Xuebao, 21, 387
  62. Liu, 2003, Molecuar cloning, Escherichia coli expression and genomic organization of squalene synthase gene from Artemisia annua, Acta Bot. Sin., 45, 608
  63. Lu, 2005, Terpenoid metabolism in cotton (Gossypium spp.) and qinghao, Adv. Plant Physiol., 8, 265
  64. Bharel, 1996, Structure, biosynthesis and functions of artemisinin, Fitoterapia, 67, 387
  65. Li, 1994, Synthesis of [15-14C] labeled artemisinin, Yaoxue Xuebao (Acta Pharmaceutica Sinica), 29, 713
  66. Nair, 1992, Use of cell-free systems in the production of the potent antimalarial, artemisinin, Indian J. Chem., Sect. B, 31B, 880
  67. Zhang, 2003, Effect of horseradish peroxidase on the biosynthesis of artemisinin in Artemisia annua in vitro, Yingyong Yu Huanjung Shengwu Xuebao, 9, 616
  68. Huang, 1990, Biosynthesis of arteannuin. I. In vivo biosynthesis of arteannuic acid of Artemisia annua, Huaxue Xuebao, 48, 275
  69. Abdin, 2004, Molecular approaches to enhance artemisinin content in Artemisia annua L., Recent Prog. Med. Plants, 4, 145
  70. Liu, 1999, Advances in artemisinin research, Huaxue Jinzhan, 11, 41
  71. Gulati, 1996, Experimental studies on Artemisia, a herbal remedy to malaria, Fitoterapia, 67, 403
  72. Chen, 1986, Factors affecting the content of artemisinin in Artemisia annua, Zhongyao Tongbao, 11, 393
  73. Chen, 1987, Studies of several physiological factors on artemisinin synthesis in Artemisia annua, Zhiwu Shenglixue Tongxun, 5, 26
  74. Chen, 1993, Enhanced synthesis of the natural phytotoxin artemisinin in tissue culture, PGRSA Q., 21, 151
  75. Zhang, 2004, Related factors of artemisinin biosynthesis in clone strain of Artemisia annua L., Yingyong Yu Huanjung Shengwu Xuebao, 10, 277
  76. Li, 1999, Artificial regulation of artemisinin biosynthesis metabolite in cultured cells of Artemisia annua, Zhongguo Shengwu Huaxue Yu Fenzi Shengwu Xuebao, 15, 479
  77. Wang, 2006, Preparation of an elicitor from a fungal endophyte to enhance artemisinin production in hairy root culture of Artemisia annua L., Shengwu Gongcheng Xuebao, 22, 829
  78. Liu, 2002, Leptosphaerone, a metabolite with a novel skeleton from Leptosphaeria sp. IV403, an endophytic fungus in Artemisia annua, Helv. Chim. Acta, 85, 2664, 10.1002/1522-2675(200209)85:9<2664::aid-hlca2664>3.0.co;2-r · DOI
  79. Huang, 2006, Genetic manipulation on biosynthesis of terpenoids, Zhongguo Shengwu, 26, 60
  80. Huang, 2008, Cloning and expression of Artemisia annua L. amorpha-4,11-diene synthase gene, Guangzhou Zhongyiyao Daxue Xuebao, 25, 68
  81. Keasling, J.D. (2005, January March). Engineering microbes for anti-malarial production. Abstracts of Papers; In Proceedings of 229th ACS National Meeting, San Diego, CA, USA.

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Metadata

Authors
Brown, G. D.
Affiliations
University of Reading (GB)
Corresponding author
Brown, Geoffrey D.
Venue
Molecules
Publisher
MDPI AG
Published
2010-10-28
Volume
15
Issue
11
Pages
7603-7698
ISSN
1420-3049 (ISSN-L)
OpenAlex
W1981404580
Cited by
363
References
328
Citations by year
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FWCI
1.46
Type
journal article
Language
en
Open access
Locations
  • Molecules (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • CentAUR (University of Reading) (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
Biotechnology and Biological Sciences Research Council (BB/G008744/1)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant biochemistry and biosynthesis
  • Physical Sciences › Chemistry › Organic Chemistry › Sesquiterpenes and Asteraceae Studies
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Drug Discovery › Phytochemical compounds biological activities
Keywords
Artemisia annua, sesquiterpenes, dihydroartemisinic acid, allylic hydroperoxides, endoperoxides, terpene oxidation, cadinane sesquiterpenes, flavonoids, stable isotope labeling, feeding experiments, antimalarial natural products, phytochemistry
MeSH terms
Antimalarials, Plants, Medicinal, Sesquiterpenes, Artemisia annua, Artemisinins

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

A language model drew the claims from the paper on 2026-10-09, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).

A second model labelled each claim on 2026-10-09 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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