Frontiers in Natural Product Chemistry: Volume 4

Plant-Based Antiamoebic Natural Products: Literature Review and Recent Developments

Neelam Bharti¹, *, Prabhu P. Mohapatra², Shailendra Singh³

¹ Mellon Institute Library, Carnegie Mellon University, Pittsburgh, PA 15213, USA

² Department of Chemistry, Brooklyn College, Brooklyn, NY 11210, USA

³ Environmental Health and Safety, Carnegie Mellon University, Pittsburgh, PA 15213, USA

Abstract

Amoebiasis, caused by Entamoeba histolytica (E. histolytica), is the third leading cause of health problems in developing countries and affects more than 10% of the world’s population. If left untreated, amoebiasis causes severe complications including hepatic and intestinal tissue destruction. According to World Health Organization (WHO), approximately 80% of the people in less-developed countries still rely on traditional medicine for their healthcare needs. In different parts of the world, plant extracts obtained from numerous plants have been used in the indigenous system of medicine for the treatment of dysentery. To apply these remedies to the rational scientific use of herbal medicines, studies on antiamoebic drugs mainly focused on natural products associated with folklore medicine. Plant extracts have been tested, and several natural products have been isolated from the active fractions, leading to the characterization and extensive biological studies of the isolated compounds. This book chapter reviews the noticeable crude extracts from medicinal plants tested for antiamoebic activities and isolated natural products that have been evaluated for antiamoebic activities in the last five decades. Most of these compounds with known structures belong to alkaloids, terpenoids, quassinoids, flavonoids, iridoids, and other phenolic compounds obtained from higher plants. These natural products showed activity against E. histolytica and other protozoa in vitro, and some of the natural products have been tested in vivo. Only a very few of them have been tested clinically. Herein, we report the traditional user’s knowledge, pharmacological activity, mechanism of action, toxicity, and other properties of antiamoebic natural products.

Keywords: Amoebiasis, Antiamoebic, Alkaloids, E. histolytica, Flavonoids, Iridoids, Natural products, Quassinoids, Terpenoids.

* Corresponding author Neelam Bharti: Mellon Institute Library, Carnegie Mellon University, Pittsburgh, PA 15213, USA; Tel: 412-268-6107; E-mail: nbharti@andrew.cmu.edu

INTRODUCTION AND BACKGROUND

Parasitic infections are the most prevalent disease throughout the world and one

of the most predominant health concerns in tropics and subtropics. Most of these infections occur due to the consumption of contaminated food or water and the lack of sanitation resulting in severe diarrhoeal condition. Diarrhoeal diseases are the second leading cause of death in children under five years, killing around 525,000 children annually [1, 2].

The World Health Organization (WHO) has estimated about 1.7 billion reported cases of diarrhoeal diseases every year, resulting in thousands of deaths annually around the world [2, 3]. Among the patients with infectious diarrhea, bloody diarrhea (amoebiasis) is one of the three major clinical conditions, causing life-threatening dysentery. Amoebiasis, caused by a protozoan parasite, Entamoeba histolytica (E. histolytica), remains a major health problem in the developing countries [4]. It affects one-tenth (10%) of the world’s population, and if left untreated, the infection can lead to severe complications including hepatic amoebiasis, liver abscess, bloody diarrhea, and intestinal tissue damage [5]. Globally, amoebiasis accounts for over 50 million cases and is responsible for ~110,000 deaths annually, surpassed by only malaria and schistosomiasis in the mortality of parasitic diseases [6-10]. The prevalence of amoebiasis is more closely associated with sanitation and poor lifestyle rather than the location and climate of the region and is preventable with proper sanitation, awareness, and treatment.

Organism: Several species of Entamoeba protozoa have been reported (Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, Entamoeba polecki, Entamoeba coli, and Entamoeba hartmanni) but only a few of them are pathogenic and associated with the disease. E. histolytica is a pathogenic protozoan which secrets enzyme proteinases. Proteinases kill host cells by dissolving the cell wall and consuming red blood cells, resulting in colitis and bloody diarrhea. Pathogenic trophozoites travel through the compromised mucosal barrier to reach liver, causing liver abscesses [4].

E. histolytica has a simple two-phase life cycle, infectious immobile cyst, and pathogenic motile trophozoite (Fig. 1) [11]. E. histolytica cysts are spherical, quadrinucleate, and enclosed by a refractive coating of chitin. Usually, the infection is initiated by the ingestion of cysts through the consumption of contaminated food or water [4, 9]. Cysts move through the digestive system, survive the stomach acid, and finally reach the small intestine and colon to excyst and form trophozoites. E. histolytica trophozoites are pleomorphic in shape and highly motile; they can ingest bacteria and food particles. Trophozoites reproduce by binary fission, encyst within the colon, and excrete into the environment with stool. Trophozoites may be present in the stool as well, but unlike the cyst, they cannot survive outside the human host [4, 9].

Fig. (1))

E. histolytica life cycle [12].

Treatment: Choice of drug for amoebic treatment depends upon the desired clinical target and site of action. The currently used antiamoebic drugs fall into two classes: tissue amoebicides and luminal amoebicides (Fig. 2). Tissue amoebicides such as metronidazole (1), tinidazole (2), and secnidazole (3) kill amoeba in the host tissue and organ, whereas luminal amoebicides such as iodoquinol (4), diloxanide furoate (5), and paromomycin (6) are active only in the intestinal lumen and absorbed poorly [9].

Fig. (2))

Amoebicidal agents in use.

Currently, metronidazole [1-(2-hydroxyethyl)-2-methyl-5-nitroimidazole] (1) and tinidazole (2) are the most effective antiamoebic drugs (IC50 = 0.018-0.032 µg/mL or IC50 = 8.2-3.2 µm, respectively); however, in the case of severe infection, it is recommended to be administered with other antimicrobial drugs [12-14]. Metronidazole (nitroimidazole) is a prodrug; ferredoxin nitroreductase enzyme reduces the nitro group of metronidazole to form an active drug molecule. Although metronidazole is the drug of choice, it has several severe side effects. It is mutagenic in bacteria and carcinogenic in rodents. Other side effects include gastrointestinal disturbance, nausea, vomiting, diarrhea, headache, and stomatitis [15-17]. The long-term systemic treatment with metronidazole is associated with the development of leucopenia, neutropenia, and/or central nervous system (CNS) toxicity [9, 18-21]. The development of resistance to metronidazole in protozoan (E. histolytica) has also been reported [22-24]. Because of the undesired side effects and development of possible resistant strains of amoeba against metronidazole, new more effective and safer amoebicidal agents are in demand [9].

Search for Herbal Antiamoebic Compounds

Medicinal herbs and herbal extracts have been a part of the traditional medicines for centuries and always been an important source of potential chemotherapeutic agents. Natural products play a significant role in the drug discovery and development process. Most modern pharmaceutical agents are based on natural products. Evidence shows that Ancient Egyptians practiced medicine in 2900 BC. The first documented use of a natural product as medicine was as far back as 78 AD [25]. Since then, different indigenous plants are in practice in various cultures around the world; they contain diverse compounds that exhibit various pharmacological and medicinal activities. More than half of the pharmaceuticals drugs in clinical use today are derivatives of natural products including several antibiotics, anticancer drug paclitaxel, and vitamin A [25]. Thirteen natural product-derived drugs were approved between 2005-2007 [26].

Before synthetic drugs were developed, diarrheal infections were treated using folk medicine based on the belief and knowledge passed down through the generations. Even now, because of the unavailability of modern medical facilities, people commonly search for natural remedies from traditional plants found in folklore. Millions of people in the third world countries practice herbal medicines and regard them as their system of medicine because of low cost, easy access, and ancestral experience [27]. According to WHO, almost 20,000 medicinal plants exist in different continents and approximately 80% of people rely almost exclusively on traditional medicines for their primary health care needs [28-30].

The development of a potential natural product as a drug candidate requires the screening of large numbers of plant extracts, isolation, and identification of active compounds, elucidating their mechanism of action, as well as toxicity tests to demonstrate selectivity towards the host and parasite. To discover active drug constituents from plants, high-throughput screening tests are in practice for bioassay-guided fractionation, leading to the isolation of active ingredients from active plant extracts [29-31]. The active fractions were developed into clinical agents either as the natural product or as a synthetic analog with enhanced clinical action or reduced adverse side effects.

Liquid concentrates or powdered plant parts are used in traditional medicine, for treatment. The concentration of the active ingredient in a plant extract depends on the region, time of harvest, and climatic conditions for growing. In different studies, plant extracts were tested, and the most active fractions underwent extensive biological and phytochemical studies. This led to the isolation and characterization of their active ingredients [29-31].

Standard Extraction Procedure

Extraction of the plant material is an important step in the analysis of medicinal plants and isolation of pharmaceutically active ingredient. Different methods are in use for plant extraction; a general procedure is described in Fig. (3).

Fig. (3))

Key steps in plant extraction for medicinal purpose.

In short, collected plants are generally identified by an expert; it is recommended to keep one specimen of the plant. The plant parts are first dried and then extracted with different solvents. Extraction of compounds depends on the polarity of solvent used for extraction. The extracts are concentrated (freeze-dried/lyophilized) at a low temperature and tested against the causative agent. Active extracts are further fractionated and tested. Purification of the active fractions affords the active ingredients that are retested and characterized [31, 32].

In the past few decades, studies in the search for antiamoebic agents from natural products led to the isolation of many new bioactive compounds. Herein, we present a compilation of such studies and highly active isolated natural products evaluated for their antiamoebic activity.

Alkaloids

Alkaloids are a class of naturally occurring nitrogen-containing compounds with pronounced physiological actions on living organisms. Examples of some highly bioactive alkaloids are morphine (7), atropine (8), and quinine (9) [33] (Fig. 4). Alkaloids have been used as medicines for humans and animals as well as poison for execution and war, as described by Mann as Murder, Magic, and Medicine [34]. The 19th century has witnessed the isolation of several bioactive alkaloids such as emetine, atropine, quinine, and caffeine (1817-1820) [35]. Later, pharmacological applications of these alkaloids were explored; they were found to be effective against various infections [36-38] (Figs. 4-11, Table 1).

Fig. (4))

Structures of morphine (7), atropine (8), and quinine (9).

Cinchona succiruba bark is well known for the isolation of quinine (9), a quinolone alkaloid. First discovered in the 17th century, quinine was used to treat common infections. Later, it was used as an antimalarial compound for more than a century during the 1600s and as one of the compounds to treat amoebic dysentery [39-41]. Several cinchona alkaloids were tested, but only a few of them showed antiamoebic properties. Quinidinone (10) is an analog of quinine prepared by the oxidation of the hydroxyl group at C-9 to a ketone; it showed more pronounced activity (IC50 = 7.4 µg/mL) than quinine. It was the most active, followed by quinine (9), and quinidine (11). Other cinchona alkaloids studied by Keene et al., 10-methoxycinchonamine (12) and aricine (13) (isolated from Rauwolfia ligustrina) were highly active against E. histolytica [42] (Fig. 5).

Fig. (5))

Structures of quinidinone (10), quinidine (11), 10-methoxy cinchonamine (12), and aricine (13).

Emetine (14), an isoquinoline alkaloid, was isolated from the root and rhizome of Cephaelis ipecacuanha in 1817; it was one of the first few drugs used against parasitic infections [42, 43]. Emetine (14) is an active ingredient in ipecac syrup and has been used in the past in many acute oral poisonings. Emetine is highly active against amoebiasis and kills the trophozoite form, but not very active against amoebic cyst (IC50 = 0.07 µg/mL). Despite the high activity, the clinical use of emetine is limited because of its severe side effects and toxicity [43]. Emetine is a known myotoxic and cardiotoxic agent. A death case was reported due to cardiac failure following the use of emetine hydrochloride [44]. The cellular mechanism of action for emetine involves the inhibition of protein and DNA synthesis in a cell and accounted for the induction of programmed cell death of E. histolytica trophozoites [45]. Emetine inhibits protein synthesis by interfering with ribosomal translocation along the mRNA by acting on 40-s ribosomal subunit. Although, regardless of its toxicity, it was used to treat severe ulcerative amoebiasis [45]. A synthetic analog of emetine, 2,3-dehydroemetine, was preferred over emetine because of its low toxicity profile, faster metabolism, and excretion rate [46, 47]. High activities of emetine and 2, 3-dehydroemetine against E. histolytica are associated with the natural configuration of the alkaloids because the removal of 9,10-dimethoxy moiety from emetine caused 52-fold loss of activity, proving the necessity of this group for amoebicidal activity. Keene et al. investigated the antiamoebic activities of 18 structurally similar alkaloids isolated from different plants [48]. These alkaloids contain either two benzyltetrahydroisoquinolines, two tetrahydro-β-carbolines, or a mixture of benzyltetrahydroisoquinoline/tetrahydro-β-carboline moieties; they were structurally very similar to emetine (14). Tubulosine (15) and its analog pseudotubulosine are significantly less active than emetine despite the same stereochemistry as emetine. An isoquinoline alkaloid strychnofoline (16) isolated from Strychnos usambarensis sharing a similar stereochemistry as emetine was inactive against E. histolytica [49]. Benzylisoquinoline alkaloid, berberine (17), isolated from Berberis aristata was found to be active against E. histolytica, both in vitro and in vivo [50, 51]. Several quinolizidine alkaloids including matrine (18), its N-oxide, and cytisine (19) isolated from a species of Sophora exhibited amoebicidal activity [52] (Fig. 6).

Fig. (6))

Structures of emetine (14), tubulosine (15), strychnofoline (16), berberine (17), matrine (18), and cytisine (19).

Apocynaceae is a flower bearing plant family that has been used for centuries as a traditional medicine in India, South East Asia, and Africa to treat dysentery, bacterial, and viral infections [53]. They are a rich source of indole alkaloids and possess the active ingredient, alstonine (20), however it was not found active

Fig. (7))

Structures of alstonine (20), alstonerine (21), pleiocarpamine (22), macrocarpamine (23), villastonine (24), and chonemorphine (25).

when tested against E. histolytica [54]. Wright et al. isolated several alkaloids from the roots of Alstonia angustifolia, including some monomeric and dimeric alkaloids. The monomeric alkaloids were alstonerine (21) and pleiocarpamine (22), while the dimeric alkaloids contain two different monomeric alkaloidal units [55]. The antiprotozoal activities of two dimeric alkaloids, macrocarpamine (23) and villastonine (24), were significant but 4–8 fold less than emetine against E. histolytica [55]. Macrocarpamine and villastonine consist of one alstonerine molecule and one pleiocarpamine molecule, even though separate studies of monomeric alkaloids, alstonerine (21) and pleiocarpamine (22), did not show any significant activity, indicating the synergistic effect of both the molecules (Fig. 7). Chatterjee et al. studied Chonemorphine (25), a steroidal alkaloid isolated from Chonemorpha fragrans for its antiamoebic properties. It cured the hepatic infection in golden hamster at a dosage of 100 mg/kg X 4, and intestinal infection in rats at 200mg/kg (X 4) [56].

The most potent alkaloid found in Wright’s study was villastonine (24); its selective cytotoxicity against amoeba and KB cells (human epidermoid carcinoma of the nasopharynx) was also investigated. It was toxic to all the cells and not selective towards protozoa or KB cells. Activity profile studies suggested that the presence of ring systems was necessary for the dimer bioactivity. Presence of the ester group suggested significantly increased lipophilicity and probably enabling its transport across lipid barriers and cell membranes.

Wright et al. also studied other monomeric alkaloids, alstophylline (26), 11-methoxyakuammicine (27), norfluorocurarine (28), and vincamajine (29), they were considerably less active than their dimers (Fig. 8) [55].

Fig. (8))

Structures of alstophylline (26), 11-methoxy akuammicine (27), norfluorocurarine (28), and vincamajine (29).

Conessine (30), conessidine (31), and conessimine (32) are the major steroidal alkaloids isolated from Holarrhena antidysenterica; the plant introduced in Europe for the treatment of dysentery in the 19th century [57] (Fig. 9). Crude extracts of tabernaemontana species were reported as potential antiamoebic extracts, and the active compounds were bisbenzylisoquinoline alkaloids containing two isoquinoline moieties linked to two benzyl moieties [58]. They were classified into 26 structural classes based on the number, position, and type of connecting bridges between the two monomers.

Fig. (9))

Structures of conessine (30), conessidine (31), and conessimine (32).

Seven different alkaloids isolated from Strychnos usambarensis were studied by Wright et al. [59]. These alkaloids were tested against E. histolytica and Plasmodium falciparum; they showed different activity profiles towards both the protozoa. Out of the seven alkaloids, usambarensine (33), 3′,4′-dihydrousambarensine (34), usambarine (35), and 18′,19′-dihydrousambarine (36) showed significant in vitro activities against E. histolytica (Fig. 10). It was proposed that different substitutions in their structure induced a positive effect on the action against one organism and a negative effect on the other organism. Usambarine (35) was found to be highly active in vitro against E. histolytica compared to P. falciparum; however, usambarine analog (strychnopentamine 37) containing C-11 hydroxy and C-12 N-methylpyrrolidine moieties showed a low antiamoebic activity [59].

These substituents at the C-11 and C-12 positions clearly affect the antiprotozoal activity profiles of the compounds, confirming again that even a minor structural difference affects the antiamoebic activity. Interestingly enough, individual alkaloids showed subtle differences for the substitution at specific sites of these compounds, which was somehow relevant to the antiamoebic activity. Molecular conformation studies showed that usambarensine may adopt a similar conformation as emetine and possibly work as a protein synthesis inhibitor similar to emetine. The mode of action of usambarine and related alkaloids is not fully known, but it is assumed that they inhibit protein synthesis, induce apoptosis, and act as DNA intercalators [60]. Emetine (14) and usambarensine (33) are related alkaloids and contain two aromatic systems in a certain arrangement, indicating the minimum energy conformation state for their activity. Cytotoxicity studies showed that usambarensine (33) and usambarine (35) are most selective against KB cells. The cytotoxicity-to-antiamoebic ratio for usambarine was 20.4, whereas, for usambarensine, this ratio was 18.8. Emetine was more toxic to KB cells than amoebae (cytotoxic/antiamoebic ratio 0.14).

Fig. (10))

Structures of usambarensine (33), 3′,4′-dihydrousambarensine (34), usambarine (35), 18′,19′-dihydrousambarine (36), strychnopentamine (37), and isostrychnopentamine (38).

Basic structural similarities of strychnopentamine (37) and isostrychnopentamine (38) to emetine made them potential antiamoebic candidates; they exhibited potent activity against E. histolytica. Compounds containing only one indole ring system did not show any activity, indicating that the presence of two indole moieties is essential for their activity [60]. An oxindole alkaloid, strychnofoline isolated from S. usambarensis, falls under inactive compounds category, probably because of the absence of two indole moieties.

Marshall et al. studied the antiplasmodial, antiamoebic, and cytotoxic activities of 24 bisbenzylisoquinoline alkaloids isolated from different plant species [61]. Emetine was used as a standard drug for comparison (IC50 = 2.23 µM). The most active alkaloids against E. histolytica in this study were aromoline (39), isotrilobine (40), and insularine (41) with IC50 values in the range 5–11 µM, whereas 19 alkaloids were active against P. falciparum with IC50 values <10 µM (Fig. 11). The activity of the tested alkaloids against E. histolytica was significantly different from that against P. falciparum but did not show significant cytotoxicity against KB cells. Another bisisoquinoline alkaloid cryptopleurine (42) isolated from Cryptolepis sanguinolenta was highly active against E. histolytica [62] (Fig. 11). It inhibits protein synthesis at a low concentration and blocks peptide translocation at t-RNA to inhibit peptide bond formation [63]. Bhutani et al. studied 14 phenanthroindolizidine alkaloids isolated from two species of Tylophora plants as antiamoebic agents [64]. All the tested alkaloids varied in number, nature, and distribution of oxygen-bearing substituents and were not significantly active against E. histolytica [64]. Hundreds of plant extracts have been tested for antiamoebic activities, and the active ingredients were isolated, Table 1 shows the notable alkaloids with their antiamoebic activities.

Fig. (11))

Structures of aromoline (39), isotrilobine (40), insularine (41), and cryptopleurine (42).

Table 1 Antiamoebic activity of isolated natural products: Alkaloids.

Terpenes and Quassinoids

Terpenes are a class of organic compounds produced by plants and animals often referred as terpenoids. Terpenes are abundant in essential oils and pigments and have been used in the treatment of parasitic infection and acute bronchitis [65-67]. Terpenes and terpenoids are the chief constituents of essential oils and are widely used as fragrances in perfumery and alternative medicines such as aromatherapy. The basic building unit of terpene is isoprene, a hydrocarbon containing five carbon atoms and eight hydrogen atoms [67] (Fig. 12).

Fig. (12))

Basic structure of terpenes [68].

Terpenes have medicinal properties such as inflammatory and cytotoxic activities [65]. Several plant extracts containing terpenes and quassinoids were found to be active against E. histolytica (Figs. 13-18, Table 2) [68, 69].

Recently, Bautista et al. explored the antiamoebic activity of sesquiterpene lactones incomptines A-D (43-46) from Decachaeta incompta [68] (Fig. 13). The antiamoebic effect of incomptines A-D (43-46) was suggested through the alteration of energy metabolism of the protozoa through the disruption of normal energy cycle mechanism [69]. Calzada et al. isolated diterpenes (polystachynes A, B, and D, as well as linearolactone) from the chia plant (Salvia polystachya) and investigated their antiprotozoal activities in vitro. Linearolactone (47) showed the most potent antiamoebic activity (IC50 = 22.9 µM) among the group, whereas polystachynes A, B, and D showed moderate antiprotozoal activity [70, 71]. A α-methylene-γ-lactone sesquiterpene, parthenin (48), isolated from Parthenium hysterophorus was also tested in vitro and in vivo against E. histolytica and cytotoxicity studies were performed on mice [72].

It showed significant in vitro activity but was less active than metronidazole when evaluated in vivo in a golden hamster model with induced liver abscesses. The hamsters were treated with four different doses; unfortunately, none of them showed the inhibition of liver necrosis compared to the reference drug. Parthenin was toxic to animals within the therapeutic range. Many natural products isolated from plant species showed activity against one or more species of protozoa, but a few are highly selective against E. histolytica [73-75]. A comparative antiprotozoal activity study of natural products indicated that the interspecies differences in protozoan protein synthesis caused the difference in selectivity towards one species or other [73].

Fig. (13))

Structures of incomptines A (43), incomptines B (44), incomptines C (45), incomptines D (46), linearolactone (47), and parthenin (48).

Quassinoids are a group of degraded triterpene lactones found in various species of traditional medicinal plants including Ailanthus, Brucea, Castela, Picrasma, Quassia, and Simarouba [75-77]. The basic structure of quassinoids is shown in Fig. (14).

Fig. (14))

Basic structure of quassinoids.

Examples of some highly bioactive quassinoids are bruceine D (49), quassin (50), and glaucarubolone (51) (Fig. 15). The first two quassinoids isolated and characterized were quassin (50) and neoquassin (52). These two compounds were isolated by Clark’s group in the early 1930s; however, their structures were not characterized until the 1960s [78]. Studies on quassinoids were extended with the isolation, structure elucidation, and pharmaceutical evaluation. Quassinoids exhibit anti-inflammatory, antiviral, antimalarial and antitumor activity [79]. Quassinoids are known as potential protein synthesis inhibitors in parasites and mammalian cells. Mechanism of action is not fully known, but it is suggested that some quassinoids affect the HI-1a protein, inhibit protein cycle, and polarize the membrane [79].

Fig. (15))

Structures of bruceine D (49), quassin (50), glaucarubolone (51), and neoquassin (52).

Several quassinoids have been used in indigenous medicine for the treatment of malaria and amoebic dysentery [80-84]. Quassinoids isolated from Brucea javanica (B. javanica), Quassia amara (Q. amara), and Simarouba glauca (S. glauca) were studied in various parts of the world against parasitic infections. Wright et al. investigated several quassinoids extracted from B. javanica fruit and S. amara stem for antiamoebic activity. Extracts of S. amara and Picrasma excelsa extracts were moderately active against E. histolytica. The main quassinoid present in the extracts was quassin (50) that was active in vivo and in vitro but was slightly cytotoxic [82]. B. javanica fruits were clinically used for the treatment of amoebic dysentery, but their extracts were less effective than emetine [82, 83]. The major constituents of B. javanica extracts were bruceantin (53), bruceine A (54), bruceine B (55), and bruceine C (56) [82, 84] (Fig. 16). Bruceine A (54) showed an IC50 of 0.097 µg/mL and was at least three times more active than bruceines B, C, and D. Bruceine A (54) has an isopropyl ester substituent, whereas bruceine B (55) has an acetate group. Bruceine C (56) contains a 3′,4′-dimethyl-4′-hydroxy-pent-2′-eneoic functionality at the C-15 position. Evidence supports that the nature of these ester groups affects the antiamoebic activity of bruceines A, B, and C. Bruceine A (IC50 = 0.097 µg/mL) is highly active followed by bruceines C (IC50 = 0.297 µg/mL) and B (IC50 = 0.306 µg/mL). Bruceantin (53) (IC50 = 0.019 µg/mL) also contains an ester group at the C-15 position, but it was many folds active than bruceines A, B, and C; and was virtually as active as metronidazole, but highly toxic to KB cells [84].

Fig. (16))

Structures of bruceantin (53), bruceine A (54), bruceine B (55), and bruceine C (56).

Glaucarubin (57) was the active molecule isolated from the extracts of S. glauca, a plant that has been used in France and Germany in traditional medicine for diarrhea treatment [85-87].

The structure–activity correlation of antiamoebic quassinoids showed that a minor structural difference in the side chain would lead to a significant difference in activity. For example, bruceantin (53) containing an isopentane sidechain is highly active followed by brusatol (58) (with an isopropyl side chain) and glaucarubinone (59) (a hydroxyl group at C-15), whereas bruceantinol (60) (five-carbon ester) is inactive. Glaucarubolone (51), glaucarubin (57), and glaucarubinone (59) showed similar activity with different side-chain structures (Fig. 17). The former had a free hydroxyl group at C-15, whereas the latter had a five-carbon ester. Glaucarubin was one of the few natural products made it to the clinical trials [87]. The drug was effective and well tolerated for the first six days at 10-280