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Chemistry

of artemisinin:

an overview*
Division of Experimental of Research, Washington, Therapeutics, Depammts DC 20307-5100, USA

H. Kyle Webster* Medicrmzl Chmiszy,

and Erich K. Lehnert Walter Reed Army Imtiture

of Parasitology and

Abstract The endoperoxide sesquiterpene lactone, artemisinin, and its derivatives have become increasingly important as antimalarial drugs with impressive activity against multidrug resistant forms of Plasmodium falciparum.Artemisinin has a novel structure among known antimalarial compounds with a unique 1,2,4-trioxane ring that is essential for activity. This paper gives an overview of the chemistry of artemisinin and comments on future prospects for artemisinin and its derivatives. Introduction Artemisinin is the name given to the active principle of qinghaosu, an extract of the Chinese medicinal plant qinghao or green artemisia (Arumisia annua L.). Qinghaosu was used for over a thousand years in China as a herbal remedy for malaria. In 1972, Chinese scientists characterized the structure of artemisinin as a sesquiterpene lactone with an internal peroxide linkage. Since artemisinin lacks a nitrogen-containing heterocyclic ring system it has a novel structure among known antimalarial compounds and its 1,2,4-trioxane ring is unique in nature (Fig. 1). The best yields (up to 0~6%) of artemisinin are obtained by petroleum ether extraction of the leaves and the flowering tops of A. annua. Other species of Artemisia do not yield appreciable amounts of artemisinin. The chemistry of artemisinin and its derivatives has been the subject of several reviews and World Health Organization (WHO) reports (WHO, 1981, 1986; XUAM-DE Luo & CHIA-CHIANG SHEN, 1987; ZAMAN & SHARMA, 1991). activity. The complete synthesis of artemisinin has been achieved by at least 3 routes. SCHMID & HOFHEINZ (1983) reported the first total synthesis of artemisinin starting from (-)-isopulegol(13 steps, 2.1% total yield). In this synthesis the 6member carbon ring of artemisinin is derived from isopulegol; alkylation of 3 side chains and functional group manipulations of these chains leads to the structure (1) (Fig. 2) which cyclizes into the desired artemisinin ring structure.

CH3

XING-XIANG ZHU et al. (1986) reported a second, longer synthesis starting from R-(+)-citronella1 (20 steps, 0.25% total yield) in China in 1983. This synthesis proceeds through the apparent natural precursor of artemisinin, artemisinic acid (Fig. 3). Ozone cleavage of the decalene ring system of artemisinic acid yields a structure similar to the intermediate used by SCHMID & HOFHEINZ (1983) for building the sesquiterpene lactone structure of artemisinin (Fig. 2).

Synthesis Artemisinin (3,6,9-uimethyl-9,10b-epi-dioxyperhydmpyranol [4,3,2-jklbenzoxepin-2.one) is a member of the amorphane subgroup of cadinenes. It is soluble in most aprotic solvents but only sparingly in oil and water. Solution in ethanol leads to degradation. Artemisinin is labile to acid or basic treatment but is stable in neutral solvents at temperatures up to 150C. Reduction of artemisinin, depending on conditions, yields desoxyartemisinin (epoxide) which is devoid of antimalarial activity or dihydroartemisinin (lactol) which is more active than the parent artemisinin. The peroxide moiety is thus essential for :.This article is nor subject IO copyright. ;gyt address: 2350 Qume Drive, San Jose, CA 95131-1807,

A third method, which has the added advantage of being selective for a single stereoisomer of artemisinin, starting from R-(+)-pulegone (10 steps, 3.6% total yield), was developed by AVERY er al. (1992). This synthesis begins with the optically active starting material and proceeds through the similar intermediate (Fig. 4). Although these methods of total synthesis show yields higher than the plant, their labour-intensive nature and

s 1128 lack of potential for scaled-up productlou make them economically unfeasible. Alternatively, the partial synthesis of artemisinin throuah another of the ~lants isolated comoounds. attcmisi& acid. is econo&csllv aopealing. &temi&tic acid exists m .%lO timed ti% concentration of artemisinin in the plant (see LAUGHLIN, 1994 and HAYES& VON~ILLER, 1994). Recenr work by ACTON & ROTH (1992) has shortened the partial synthesis to 3 steps, a reduction of the exocyclic double bond followed by II photo-oxidation (single?oxygen, 102: which proceeds through a classical ene reaction with rearrangement of the endocyclic double bond. Ring cleavage induced by air oxidation (triplet oxygen, 302) and reformation then completes the riig structure as perfotmed in most of the total synthesesof artemisinin. The 3 steps in AUON & ROTHS11992) method are simolified from the 8 steps for the t&sf&n~tion of artemi&nic acid to artetmsinin described by XINGXIANG ZHO et a/. (1986), and the total yield is increasedto more than 32% (Fig. 5).

This development, in addition to being a possible method of increasing the plant yield of attemisini+ also suggestsB possible biomechanism for the producttun of artemisinin in the plant. Arrenusinic acid and mevalonic acid lactone have been shown by labelling experiments to bc intermediates in the biosynthetic pathway; however, not much further is known about the natural biosynthesis of artemisinin. Early preparations of artemisinin (qinghaosu) tablets had poor efficacy (15% cure rate), attributed to poor absorpiion due to ihe relative ins&bility in wate;. Initial attempts to improve bioavailabiliry involved parentera formulations (ml solutions and suspensionsand aqueous suspensions). Radical cure rates increased (7C87%) wirh thesepreparations. Further study showed that the parent compounds activity and solubility could be improved by chet&cal modification, so long as the peroxide linkage was meserved (WHO. 1986). Conseouentlv, dihvdroartemisinin was &ed te prepare variouseesterd,erhe& and succinyl derivatives. Under acidic conditions derivatives were primarily pepimers, whereas under alkaline condiuons a-epimers were predominant. Differems in antimalarial activity betwe& (Yand b epimers have not been observed. Of the various dihvdroartemisinin derivalives prepared, 2 have received c&iderable attention, arremether (methyl ether derivative) and artesunate (water soluble half-ester succinate derivative). WHO (1986) rec. ommended development of another derivative, arteether (P-ethyl ether), based on its stability and formulation properties. Sodium artelinate has been studied at the Walter Reed Army Institute of Research (WRAIR), brcause of its water solubility and the efficacy of the oral formulation (LIN et al., 1987; KLAYM~~Net al., 1991; Fie 6, - %&t the complex structure and synthesis of artemisinin and its semi-synthetic derivatives, recent efforts have been directed at the synthesis of simple trioxanes that retain the 1,2,4-triosane ring. Catalytic reaction of aldehydcs (or ketones) with hydtoperoxides, 1,2-dioxetams or endoperoxides results in a variety of synthetic trioxane molecules formed when the aldehyde captures a

WR279138

0 1

s1:29 zwitterionic peroxide. Structure activity analysis reveals that the minnnum requirements for a synthetic peroxide are the 1,2,4-trioxane ring and B second common ring. Additional requirements involve an appropriate conformation, steric congestion, and chemical stability (WHO? 1986). Series of tetroxanes have also been prepared. Several synthetic trioxanes and tevoxanes have been shown to have parent antimalarial activity in vim and in viva. WR279137 and WR279138, rricyclic 1,2,4-trioxanes repared by losner and colleagues at Johns Hop F tns University. were recently shown to be as active as arteether against multi-drug resistant PIasmodium fnkiparum in Aom monkeys. These synthetic peroxides are appealing because they are relatively inexpensive and easy to produce (POSNER et al., 1992; Fig 7). Although the considerable clinical experience with artemisinin, artesunate and arremether has revealed no serious side-effect, lirtlc is known about the adminisrration of these drugs at high dosages and over long periods of time for prophylaxis. Recently Brewer and colleagues at the WRAIR reported evidence for neurotoxicity in experimental animals (see BREW?, 1994:. Their studies suggest the production of a long-hved toxic drug metabolite. The implicarion of their observatmn for the treatment of human patients needs co be determined. Perhaps it is instructive to note that the product of another species of Artemisia has achieved historical notoriety (ARNOLD, 1989). At the beginning of the 20th century absinthe? a popular liqueur, was banned world-wide because of tts devastating neurotoxicity. Absinthe was prepared by steam distillation of A. absinthiunr (wormwood). Thr active principle, thujone, whilst not structurally comparable to artemisinin, may share some of its pharmacological properties (Fig. 8).

Acron, N. & Roth, R. J. (1992). On the conversion of dihydroartemmmc acid inro anemiainin. &wmal of Organrr Chrmuty, 57,161&1614. Arnold, W. N. (IVSV). Absinthe. Sctennfic Anmum (June issue), 112-117. Avery, M. A., Chong, W. K. M. &Jennings-White, C. (1992). Stereoselective total symhesis of (+I-anemisinin, the armmalad consriruent ofhmmio nnnu~ L pour& offhe Amman ChcmiculSocte~, 114,97&979. Brewer, T. G., Peg+, J. 0 , Grace, S. J. Perras, I. M., Levme, B. S., Weina, P. J., Swearengen, ]., Heiffer, M. H. & Schuster, B. G. (1994). Neurotoxicirv in animals due to arreerher and artemether. Tronsacrionr D de Royal So&y of Tropical Medicine and Hygwze, 88, supp f emend 1,33-36. Havnes, R. K. & Vonwiller, S. C. (1994). Extractiop oiartemisi&n and artemisinic acid: preparation of artemether and new andogues.Tranracrias of rhe Royal Somy of Tmpd MrdmeondHygm+ 88, su plemcnt 1.2>26. Klayman, D. L., Ager, A. E ., ir, Neckensrein, L. & Lm, A. J. (1991). Tmnsdermal arteliic acid: an effecuve treatmenr for Plarmodtum bergher-infected mice. .4mrricaa ~oumol of Tmptul.Usdicine and Hygiene, 45,602-607. Laughlin, J. C. (1993). Agricultural production of artemisinina review. Tronsocrms of the Royal Sooetv oJTrop~cal Medmw andHygme,88, supplement I, 21-22. Posner, G. H., Oh, C. H., Gerena, L. & Milhous, W. K. (1992). Extraordinarily potent anrimalar,al compounds: new, structurally simple, easily synthesized, tricyclic l,Z,Ctrioaarm ~ouml o/MedrcinolChemirtq, 35,245%2467. Schmld, G. & Hofheim, W. (1983~. Total synthesis of qmghgh2j,Jmmal of t/u Ammcon Chrmml Soaq, 105, WHO (198 I). Reporr of thefourrhmemng thpsimttfic vorhmg of group on the chen,orhwa~.v of Melanie dmelopmmr of ad IU demo&s es oncime&ial drags. Geneva: War pmfihooru d ealtb Organization, rmmeogmphed documenr no. TDPJChemalSWG/QHSSl-3. WHO (19861. Reporr of a neerrng of the simtnJic workinggroup on rha ckmotherapy of malaria: the dmxbpwmt of ortmisinin md us denvnrivas. Geneva: World Health Organizauon, mimeographed docunenr no. TDR/Chemal-SWG/ART/86 3. Xing-Xieng Xu, Jx Zhou, Da-Zhang Huang & Wei Shong Zhou :1986). Total synthesis of artemisinin and deowarrcmsinin. Tetrahedron? 42,819-828. Xuam-De Luo & Chia-Chiang Shen (1957). The chermsrry, pharmacology and clinical applicatmns of qinghaosu (artcmisinin) and itsdenvatives. Med~calRaseorchReum, 7,29-52. Zaman, S & Sharma, R. P. (1991). Someaspects of the chemtrtry and bIologica activity of arremisinin and rrlared anrimalarials. Hmrocyrler, 32, 1593-1638.