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Anais da Academia Brasileira de Ciências (2011) 83(4): 1373-1384 (Annals of the Brazilian Academy of Sciences) Printed version ISSN 0001-3765 / Online version ISSN 1678-2690 www.scielo.br/aabc
Folk uses and pharmacological properties of Casearia sylvestris: a medicinal review
PAULO MICHEL P. FERREIRA1, LETÍCIA V. COSTA-LOTUFO2, MANOEL O. MORAES2, FRANCISCO W.A. BARROS2, ALINE M.A. MARTINS2, ALBERTO J. CAVALHEIRO3, VANDERLAN S. BOLZANI3, ANDRÉ G. SANTOS4 and CLÁUDIA PESSOA2 1Departamento de Ciências Biológicas, Campus Senador Helvídio Nunes de Barros, Universidade Federal do Piauí, Rua Cícero Eduardo, 905, 64600-000 Picos, PI, Brasil 2Departamento de Fisiologia e Farmacologia, Faculdade de Medicina, Universidade Federal do Ceará, Rua Cel. Nunes de Melo, 1127, 60430-270 Fortaleza, CE, Brasil 3Núcleo de Bioensaios, Biossíntese e Ecofisiologia de Produtos Naturais, Instituto de Química, Universidade Estadual Paulista, Rua Francisco Degni, s/n, 14800-900 Araraquara, SP, Brasil 4Laboratório de Farmacognosia, Departamento de Princípios Ativos Naturais e Toxicologia, Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista, Rodovia Araraquara, Jaú, Km 1, 14801-902 Araraquara, SP, Brasil
Manuscript received on December 21, 2009; accepted for publication on May 19, 2011
ABSTRACT Folk uses and scientific investigations have highlighted the importance of Casearia sylvestris extracts and their relevant bioactive potential. The aim of this work was to review the pharmacological properties of C. sylvestris, emphasizing its anti-ulcer, anti-inflammatory, anti-ophidian and antitumor potentialities. Ethanolic extracts and essential oil of their leaves have antiulcerogenic activity and reduce gastric volume without altering the stomach pH, which corroborates their consumption on gastrointestinal disorders. Leaf water extracts show phospholipase A2 inhibitory activity that prevents damage effects on the muscular tissue after toxin inoculation. This antiphospho- lipasic action is probably related to the use as an anti-inflammatory, proposing a pharmacological blockage simi- lar to that obtained with non-steroidal anti-inflammatory drugs on arachidonic acid and cyclooxygenase pathways. Bioguided-assay fractionations lead to the identification of secondary metabolites, especially the clerodane diterpenes casearins (A-X) and casearvestrins (A-C), compounds with a remarkable cytotoxic and antitumor action. Therefore, the C. sylvestris shrub holds a known worldwide pharmacological arsenal by its extensive folk utilization, exciting searches for new molecules and a better comprehension about biological properties. Key words: Casearia sylvestris, folk uses, pharmacological properties.
INTRODUCTION nas and Kinghorn 2005, Buriol et al. 2009). Although there is a great interest in synthetic techniques, such as The phytotherapic use has increased in developing and combinatorial chemistry, natural product prospection is in developed countries as an additional way to treat and a continuous source of new drugs, and around 48% of prevent diseases, particularly chronic illnesses, such new chemical entities (NCE) from 1981 to 2002 were as cardiovascular and neurodegenerative dysfunctions natural products or natural product-derived (Newman (World Health Organization 2003, Ferreira et al. 2008). et al. 2003). In spite of the medicinal importance, the These natural products are represented by teas, powders, toxicological potential in plants is frequently considered extracts, infusions and other sorts of formulations (Balu- superfluous and dealt with low priority (Junior and Correspondence to: Cláudia Pessoa Pinto 2005, Ferreira et al. 2007). E-mail: [email protected]
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The Brazilian territory embrace the major diversity The flowers are small, green, actinomorphic, hy- of plant species in the world, but less than 10% have pogene and hermaphrodite. They produce impercepti- been evaluated with respect to their biological character- ble odor, nectar of easy access and have exposed sexual istics and fewer than 5% have been subjected to detailed organs. The plant flourishes in June and August and its phytochemical studies (Luna et al. 2005). Nowadays, fruits ripen from September to November. These fruits the curative principle of plants is more disseminated in are septicidal capsules with seeds involved by red aril rural areas due to the lack of efficient laboratorial and (Absy and Scavone 1973, Torres and Yamamoto 1986). clinical facilities, where medical services are problem- Under good conditions, C. sylvestris is capable to flower atical, complex and financially more expensive (Holetz and fruit in the second year of life, when it reaches et al. 2002, Ferreira et al. 2009). about 1 meter in stature (Little and Wadsworth 1964). In Panamá, these species flower even in dry season be- TAXONOMYOF Casearia sylvestris tween August and December (Croat 1978). With a mod- Traditionally, Casearia sylvestris Swartz, 1860 (Syn- erate rate of growth, the plant lives at least 20 years. Its onymous: Samyda parviflora L., Casearia parviflora L., natural and artificial propagation usually occurs through Anavinga samyda Gaertn. F.) belongs to the Flacour- seeds and its wood is used as fuel and suitable to build tiaceae family, which has 89 genus and 1300 species fence, posts, stakes, rustic carpentry and handle tools distributed in tropical and temperate regions around (Torres and Yamamoto 1986). the world. Casearia genus includes almost 180 species Acquainted with the medical importance of several (Little and Wadsworth 1964, Torres and Yamamoto parts of Casearia sylvestris, this work aims to review its 1986). Recently, molecular, ecological, chemical and folk pharmacological uses, emphasizing its anti-ulcer, morphological studies associated with phylogenetic ap- anti-inflammatory, anti-ophidian and antitumor poten- proaches of several families, including Flacourticaceae tialities. and Salicaceae, led to the birth of the Angiosperm HARMACOLOGICAL ROPERTIES Phylogeny Group (APG), leading to the Flacourtiaceae P P family extinction and permutation of the majority of In Brazil, C. sylvestris is popularly known as “guaça- its genus to the Salicaceae family, including Casearia tonga”, a word originated from “Tupi-Guarani” lan- (Souza and Lorenzi 2005). guage, which indicates its ancient exploitation by native communities. Other folk names are “erva-de-lagarto”, DISTRIBUTIONAND ANATOMIC DESCRIPTIONOF “língua-de-tiú”, “cafezinho-do-mato”, “corta-lengua”, Casearia sylvestris bugre herb and wild-coffee (Hoehne 1939, Little and Commonly widespread in the Americas, C. sylvestris is Wadsworth 1964). The Karajá Indian tribe habitually found in Cuba, Antilleans and Porto Rico to Mexico, makes a bark maceration to treat diarrhea; other Brazil- Brazil, Bolivia, Peru and Argentina (Little and Wads- ian tribes crush its roots to care for wounds, leprosy worth 1964, Instituto de Botanica Darwinion 2002). In and as anti-ophidian. Natives from the Shipibo-Conibo Brazil, it is distributed practically throughout the en- tribe (Peru) utilize bark decoction for flu and chest tire territory extension from Amazonas (Tapajós river colds (Hoehne 1939). Certainly, the antimicrobial region) to Rio Grande do Sul states (Torres and Ya- action against pathogens as fungi (Aspergilus niger, mamoto 1986, Hack et al. 2005). Saccharomyces cerevisiae, Candida albicans, Candida Anatomically, its vegetative and reproductive tropicalis) (Carvalho et al. 1998, Oberlies et al. 2002, structures are characterized by crystalline inclusions Da Silva et al. 2008b) and bacteria (Bacillus subtilis, and glandular cells full of essential oils. The leaves are Bacillus cereus, Staphyloccoccus aureus, Enterococcus assimetric, smoothes or rough, peciolated, lanceolate, faecalis, Salmonella enteriditis) (Chiappeta et al. 1983, brilliant, with 6-12 cm length and 3-5 cm wide. It pres- Mosaddik et al. 2004, Da Silva et al. 2008b) explains the ents radiate and protostelic root and the stem is siphonos- uses to treat wounds, skin ulcerations and diarrhea. telic and ectophloic, distinguished by prismatic crystals In the last years, C. sylvestris has been exploited and druses, mainly in the cortical phloem (Absy and in the development of creams against herpes and com- Scavone 1973). mercialized in mixed formulations with other plants to
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TABLE I Folk therapeutic uses of Casearia sylvestris in some countries Part of the plant/Country Traditional therapeutic use Preparation/Administration Reference Aerial part bark, Peru Diarrhea Water extract, oral Duke and Vásquez (1994) Aerial part bark, Brazil Fever Decoction, oral Brandão et al. (1985) Anti-inflammatory Decoction, oral Hirschmann and de Arias (1990) Snakebites Infusion with bark, topic Ruppelt et al. (1991) Diarrhea Maceration, oral Duke and Vásquez (1994) Leaves, Brazil Fever Decoction, oral Brandão et al. (1985) Snakebites, herpes and syphilis Decoction, oral Hirschmann and de Arias (1990) Sexual stimulant Decoction, oral Hirschmann and de Arias (1990) Sedative, tonic, Decoction, oral Simões et al. (1999) anti-inflammatory, skin diseases and diarrhea Leaves, India Snakebite Infusion Selvanayahgam et al. (1994) Leaves, Colombia Ulcers Hot water extract, topic Garcia-Barriga (1975) Various parts, Bolivia Analgesic, antacid, Numerous, oral Baya (2001) anti-inflammatory, anti-ulcer, antitumor, antiseptic, cicatrizing, depurative, haemostatic, snakebite Roots, Brazil Body pains Decoction, oral Hirschmann and de Arias (1990) Wounds, leprosy Maceration, oral Duke and Vásquez (1994) Seeds, Brazil Leprosy Oil, topic Duke and Vásquez (1994) prepare teas and infusions with therapeutical purposes causes enzyme inactivation (Constanzo 2004). Leaf (Camargo et al. 1993, Alves 2000, Simões et al. 1999). ethanolic extract do not interferes in the aminoacid ab- An infusion can be easily obtained (20 g of dried leaves sorption, displaying an additional advantage when com- per 1 L of water) for oral consumption (2-3 times/day) pared to some synthetic drugs as atropine and cime- together with meals, in case of gastric ulcers or for tidine, molecules that arouses gastric pH alterations topical treatment of wounds, burns, rashes, eczema (Torregrosa 1997). and vitiligo (Blanco 2010). C. sylvestris extracts were also capable to prevent Table I displays some therapeutic properties of early establishment of gastric lesions in stressed rats C. sylvestris and Table II shows the biological activities with efficacy similar to cimetidine and misoprostol, already mentioned by the scientific community. though these drugs have demonstrated greater pro- tection against severe lesions, especially necrotic and hemorrhagic injuries (Aboin et al. 1987). Esteves et Anti-ulcer activity al. (2005) demonstrated a decrease in gastric volumes Ethanolic extracts of C. sylvestris leaves have revealed without physiological alterations in H+ concentration, anti-ulcer properties, a finding that corroborates its wide emphasizing that essential oils as well as crude extracts use in gastrointestinal disorders (Hoehne 1939, Fialho of “guaçatonga” leaves have anti-ulcer properties. et al. 2010). Basile et al. (1990) showed that ethanolic Quantitative analyses of the ethanolic extract indic- extract significantly decreases stomach volume (42%) ated traces of terpenes (triterpenes and sesquiterpenes), in a more efficient way (when compared to misoprostol, cumarines, tannins (flavonoids) and some volatile oils 26%) and chloride acid (HCl) production in absence of that play an important role in mucosa protection (Ser- gastric pH changes, an excellent finding since protein tié et al. 2000). Flavonoids are phenolic compounds digestion is largely activated in the stomach by pepsin with antioxidant action and strong capacity to inhibit the enzymatic action. The pepsin generation from its in- production of both nitric oxide and the TNF (tumoral active precursor pepsinogen requires acid pH (between necrosis factor) mediators (Kawada et al. 1998). In 1.6-3.2) and its catalytic activity also needs acid en- fact, the antioxidant potential was showed by Menezes vironment. Therefore, alkalinization softly up to pH 5 et al. (2004) and among all bioactive compounds found
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TABLE II Biological activities from different parts of Casearia sylvestris. Part of Biological activity Extract, compound or preparation Reference the plant Leaves Antitumor Ethanolic extract, Itokawa et al. (1988, 1990) Casearins (A-F) Gallic acid derivatives Da Silva et al. (2009) In vitro cytotoxicity on normal Casearins (A-X) Morita et al. (1991), and/or tumor cells Wang et al. (2009), Ferreira et al. (2010), Santos et al. (2010) Casearvestrins A, B and C Oberlies et al. (2002) Antiviral against Herpes simplex Essential oil Da Silva et al. (2008c) Anti-inflammatory Ethanolic extract Camargo et al. (1993) Infusion Ruppelt et al. (1991) Hydroalcoholic extract Almeida (2000) Decoction 5% Sassioto et al. (2004) Essential oil Esteves et al. (2005) Antibacterial Not described Hoehne (1939), Chiappeta et al. (1983) Antifungal Methanolic extract Alves (2000) Casearins G, S and T Carvalho et al. (1998) Casearins A, B and C Oberlies et al. (2002) Phospholipase A2 inhibition Gallic acids Da Silva et al. (2008b) Water extract Borges et al. (2000, 2001), Cavalcante et al. (2007) Anti-ulcer Ellagic acid derivatives Da Silva et al. (2008a) Ethanolic extract Basile et al. (1990), Sertié et al. (2000), Fialho et al. (2010) Essential oil Esteves et al. (2005) Molluscicide Methanolic extract Alves (2000) Antioxidant Water extract Menezes et al. (2004) Against Leishmania donovani, Ethanolic and hexane extracts Mesquita et al. (2005, 2007) Trypanosoma cruzi and Plasmodium falciparum Analgesic Hydroalcoholic extract Mattos et al. (2007) Hypocholesterolemic Methanolic extract Schoenfelder et al. (2008) Stem Against L. donovani and T. cruzi Ethanolic and hexanic extracts Mesquita et al. (2005) Against Aedes aegypti Hexane extract Rodrigues et al. (2006) Against P. falciparum Hexane extract Mesquita et al. (2007) Roots Analgesic Bark infusion with leaves Ruppelt et al. (1991) Against T. cruzi Clerodane diterpen Espíndola et al. (2004) Against L. donovani, T. cruzi Ethanolic and hexane extracts Mesquita et al. (2005, 2007) and P. falciparum in the ethanolic extract of C. sylvestris, the therapeutic activity (Abreu Gonzaga et al. 2003). An enriched frac- effects of essential oils have been attributed to the bici- tion of diterpenes extracted from leaves of C. sylvestris clogermacrene, a sesquiterpene with anti-inflammatory presented antiulcerogenic action in rats in a dose of
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15.5 mg/kg/day, inhibiting ulcer formation up to 100%. ence of anti-inflammatory phytochemical compounds The clerodane diterpenes casearins B, D, O, X and and prostaglandin synthesis inhibition, which may cause caseargrewiin F (Fig. 1) presented the same action in a inability in macrophage chemotaxis, fibroblast activa- dose of 1.68 mg/kg/day. Casearin X was called initially tion and collagen fiber arrangement and formation, with as Casearin U (FAPESP et al. 2003, Santos 2008). noticeable inhibitory effects on osteogenesis (Sassioto et Besides protecting the stomach mucosa, the al. 2004). ethanolic extract shows a LD50 value of 1842 mg/kg (Basile et al. 1990), a value considered slightly toxic according to toxicity categories established for humans Anti-ophidian activity by Hodge and Sterner (1949). Some subacute toxicity In many countries, plant extracts have been tradition- assays exhibited promising outcomes, since doses 2.5 ally used in the treatment of snake bite poisoning (Martz fold higher (143.9 mg/kg/day of body weight) show ef- 1992). Water extract of C. sylvestris leaves shows great fectiveness as anti-ulcer agent without altering physio- inhibitory effects on the toxic, enzymatic, hemorrhagic logical parameters, such as water and food intake, body and anticoagulant activities exhibited by various venoms, weight gain and key organ relative weights after thirty particularly against those belonging to the PLA2 I (Phos- days of ethanolic extract oral administration (Basile et pholipase A2) (Elapidae Family), II (Viperidae Family) al. 1990). Oral acute toxicity (30, 65 and 172 mg/kg/ e III (bee) types (Borges et al. 2000, 2001). day) with the compound Casearin X also did not in- Neuromuscular junction is the main target of snake dicate corporal or behavioral changes, despite the fact venoms, some of which possessing strong pre-synaptic that 172 mg/kg/day is a dose 100-fold higher than neurotoxins, such as the Crotoxin (CTX), a heterotri- ED100 (maximum effective dose) necessary to obtain meric Asp49-PLA2 complex found in the Crotalus antiulcerogenic action (1.68 mg/kg/day) (FAPESP et durissus terrificus venom (Kini 2003). The PLA2 en- al. 2003, Santos 2008). zyme activity of CTX is critical for blockage of sig- nal transmission at the endplate. The C. sylvestris anti- ophidian property seems to be related to the capacity Anti-inflammatory activity of PLA2 inhibition (Borges et al. 2000, 2001), which is Researchers have emphasized the anti-inflammatory an enzyme largely activated after venom administration/ potential of substances extracted from “guaçatonga” inoculation. Aqueous extract of the leaves diminishes leaves, attracted by its economical appealing, practical injures on skeletal muscle fibers via the prevention of handling and easy folk preparations (Hoehne 1939, edema formation within endomise (connective tissue) Ruppelt et al. 1991). and reduces the muscle blockage, findings responsible In some carrageenan-induced inflammation by tissue damage protection and enhanced survival rates models, the essential oil of C. sylvestris leaves inhib- in extract-pretreated animals depending on the venom ited paw edema formation. This proposes a blockage of class (Cavalcante et al. 2007). inflammatory cascade mediators from the arachidonic The anti-venom activities of the extract on myo- acid and inhibition of cyclooxygenase (COX) catalytic toxins (Bothrops and Crotalus genus) is attributed to activity, a property comparable to indomethacin (a non- their antiphospholipasic action, explaining, at least par- selective and non-steroidal COX inhibitor). On the other tially, the anti-inflammatory potential in some clinical hand, this oil does not interfere on vascular permeabil- circumstances in which there are synthesis and release ity (Esteves et al. 2005) probably due to its failure in of prostaglandins and leukotrienes and failure to prevent antagonizing mastocyte and/or basophile degranulation chronic responses (Esteves et al. 2005). It could be a and discharging of active pharmacological substances plausible proposal to assume that the mechanism of (histamine and serotonin, for example) (Tamura et action of these compounds may be analogous to non- al. 2002). steroidal anti-inflammatory drugs. Decoction of C. sylvestris leaves was capable to In view of the fact that the water extract was more decrease bone formation possibly because of the pres- efficient in neutralizing hemorrhagic metalloproteinases
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15
16 14
13
12
11
H 20 R1 1 9 17 2 10 8
3 5 7 4 6 R5 19 R4 18 O R3 R2
α α OCH3 ( ) CH3CO2 CH3CO2 OH ( ) n-C3H7CO2 1.0 α α OCH3 ( ) CH3CO2 CH3CO2 CH3CO2 ( ) n-C3H7CO2 8.5 α α OH ( ) CH3CO2 CH3CO2 CH3CO2 ( ) n-C9H19CO2 0.77
OH (α) n-C3H7CO2 CH3CO2 OH (α) n-C3H7CO2 1.8 α α OH ( ) OCH2CH3 CH3CO2 OH ( ) n-C9H19CO2 4.7 α α OH ( ) OCH2CH3 CH3CO2 OH ( ) n-C3H7CO2 2.9 α OCH3 ( ) CH3CO2 CH3CO2 H n-C3H7CO2 0.17 α OH ( ) CH3CO2 CH3CO2 H n-C3H7CO2 0.37 α OH ( ) CH3CO2 n-C3H7CO2 H n-C3H7CO2 0.51 α α OCH3 ( ) n-C3H7CO2 CH3CO2 OH ( ) n-C3H7CO2 1.1 α α CH3CO2 ( ) CH3CO2 CH3CO2 OH ( ) n-C3H7CO2 0.52 α α OCH3 ( ) n-C3H7CO2 CH3CO2 CH3CO2 ( ) OH 1.6 α α OH ( ) n-C3H7CO2 n-C3H7CO2 CH3CO2 ( ) OH 1.8 α α OCH3 ( ) CH3CO2 n-C3H7CO2 CH3CO2 ( ) n-C3H7CO2 5.9 α α OCH3 ( ) n-C3H7CO2 CH3CO2 CH3CO2 ( ) n-C3H7CO2 6.0 α α OCH3 ( ) CH3CO2 CH3CO2 CH3CO2 ( ) CH3CO2 7.8 α α OH ( ) CH3CO2 CH3CO2 CH3CO2 ( ) n-C3H7CO2 4.3 α = O CH3CO2 CH3CO2 OH ( ) n-C3H7CO2 5.4 α OCH3 ( ) = O CH3CO2 H n-C3H7CO2 α β OCH3 ( ) CH3CO2 CH3CO2 CH3CO2 ( ) n-C3H7CO2 α α OCH3 ( ) OCH3 CH3CO2 OH ( ) n-C3H7CO2 α α OH ( ) OCH3 CH3CO2 OH ( ) n-C3H7CO2 α α n-C3H7CO2 ( ) n-C3H7CO2 CH3CO2 OH ( ) H β α (CH3)2CHCO2 ( ) CH3CO2 CH3CO2 OH ( ) H β α CH3CH2CH(CH3)CO2 ( ) CH3CO2 CH3CO2 OH ( ) H β α n-C5H11CO2 ( ) CH3CO2 CH3CO2 OH ( ) H α α n-C3H7CO2 ( ) CH3CO2 CH3CO2 OH ( ) H
Fig. 1 – R1-R5 substituents of casearins A-X, caseargrewiin F and casearvestrins A-C (Carvalho et al. 1998, Itokawa et al. 1990, Morita et al.
1991, Oberlies et al. 2002, Wang et al. 2009, Ferreira et al. 2010, Santos et al. 2010) and cytotoxicity (IC50) on Chinese hamster V-79 cells of casearins A-R (adapted from Morita et al. 1991).
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MEDICINAL PROPERTIES OF Casearia sylvestris 1379 from bothropic venoms than neutralize serina-proteases, al. 1991, Wang et al. 2009, Santos et al. 2010) and ca- it is proposed that this extract may act as a metal natural seargrewiin F (Santos et al. 2010), isolated from leaves; chelating (Borges et al. 2001). Other possibility would casearvestrins A-C, isolated from leaves and branches be linkage of the active compound, present in the ex- (Oberlies et al. 2002); and rel-19S-acetoxy-18R-buta- tract, to the enzyme catalytic site which could avoid noyloxy-18,19-epoxy- 6S-hidroxy-2R-(2-methylbutano- coupling of the substrate to the enzyme in a covalent yloxy) -5S, 8R, 9R, 10S-cleroda-3,13(16),14-triene, isol- or non-covalent interaction way. Highly reactive tannins ated from roots (Espíndola et al. 2004) (Fig. 1). presented in the leaves can bind to biological polymers Casearins and their biological activities show struc- and enzymes, allowing the electron transportation and ture-activity relationship largely attributed to the diter- obstructing the enzymatic activity (Abdulla and Gruber pene skeleton substitutions at positions C-2 (R1), C-6 2000, Delfino and Canniatti-Brazaca 2010). (R4), C-7 (R5), C-18 (R2) and C-19 (R3). Figure 1 de- Four ellagic acid derivatives from aqueous extracts scribes the cytotoxic activity of the casearins on Chi- of C. sylvestris were isolated, characterized and tested nese hamster V-79 cells. Casearins G, H and I, whose against actions of both total venom and PLA2 (Asp 49 structures are characterized by the oxygen-bearing func- BthTX-II) from Bothrops jararacussu venom. The iso- tion absence at C-6, showed the highest antiprolifera- lated compounds were ellagic acid, 30-O-methyl ellagic tive activity, followed by the diterpenes J, K, L and M, acid, 3,30-di-O-methyl ellagic acid and 3-O-methyl- whose structures present hydroxyl moiety at C-6 or C-7. 30,40-methylenedioxy ellagic acid. The constant inhibi- Other casearins, as N, O, P,Q and R have weaker activity, tion values for enzymatic activity as well the IC50 which seems to be determined by methoxyl, hydroxyl or values found in the edematogenic and myotoxic activ- oxygen substituents of R1. Derivatives converted from ities indicate that the ellagic acid is the best inhibitor the casearins A and D by inducement of acyl substituent of these activities. The results show that the presence at C-6 exerted decrease in the cytotoxicity (Morita et of hydroxyls at position 3 or 30 increase the capacity al. 1991), demonstrating that bulkiness at C-6 has the of these derivatives on inhibiting toxic effects. How- highest influence on the bioactivity. Such cytotoxicity ever, the existence of methoxyl groups at position 3 or on cancer cells was also found in the C. sylvestris leaf 30 reduced but did not completely inhibit the capacity oil (Da Silva et al. 2008c), whose composition previ- of compounds on reducing all the toxic effects studied ously reported exhibits two well characterized cytotoxic (Da Silva et al. 2008a). sesquiterpenes (β-caryophyllene e a-humulene) (Tatman and Mo 2002, Esteves et al. 2005, Sylvestre et al. 2005). Although the cytotoxic mechanism is unclear, Car- Antitumor activity valho et al. (1998) reported that clerodane diterpenes G, Most of the compounds isolated from Casearia leaves S and T can cause acetylating on DNA molecule. Re- are represented by a series of oxygenated tricyclic cis- cently, this toxicity on DNA molecules was confirmed clerodane diterpenes in which the tetrahydrofuran ring by Ferreira et al. (2010) who showed that Casearin X is C-acylated at positions C-18 and C-19 (Fig. 1). The causes DNA and nuclear fragmentation, phosphatidyl- diacetalic ring is a highly oxygenated structure rarely serine externalization, activation of the effector caspases seen in natural molecules, being a protected dialdehyde 3/7 and mitochondrial depolarization in leukemia cells, (Santos et al. 2007). Bioguided fractionation assays all evidences of apoptosis cell death. Caspases, cysteine- have regularly revealed that clerodane diterpenes, pre- proteases with ability to cause cleavages on laminin and dominantly those rich in oxygen atoms, possess anti- changes on DNA structure triggered by DNAses lead to fungal, antibacterial, insecticidal, antifeedant and cy- cytoskeleton and nuclear destabilization and chromatin totoxic properties (Morita et al. 1991, Merrit and Ley fragmentation and condensation. Eventually, cells be- 1992, Oberlies et al. 2002, Vieira-Junior et al. 2009). come divided in apoptotic bodies full of organelles Twenty eight clerodane diterpenes typical of and/or nuclear material surrounded by integral plasma Casearia were isolated from C. sylvestris: casearins A-X membrane. Immediately, the largest part of the bodies (Carvalho et al. 1998, Itokawa et al. 1990, Morita et is engulfed by macrophages. Thus, since there is no
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1380 PAULO MICHEL P. FERREIRA et al. intracellular factor releasing, apoptosis occurs without (Yamamoto et al. 1995). Then, the anticancer poten- immune system activation (Strasser et al. 2000). tial exhibited by MGDE and IGDE might be related Ethanolic extract phytochemical investigations of to the modulation of the immunologic system as a re- C. sylvestris leaves reported strong antitumor activity on sult of enhancement in NK cell activity in tumor-bearing Sarcoma 180 transplanted mice (100 mg/kg/dia), with mice. Therefore, many researches have highlighted the tumor growth inhibition rates higher than 80% (Itokawa in vitro and in vivo antiproliferative capacity from clero- et al. 1988). Bioguided studies led to the isolation of dane diterpenes of C. sylvestris on tumor cells, very im- six pharmacologically active compounds, the casearins portant discoveries that emphasize this plant as an im- A, B, C, D, E and F. Additional in vivo experiments portant natural source of leading molecules. (15 mg/kg/day) confirmed the antineoplasic action of Besides this antitumor potentiality, different parts these clerodanes, though casearin C has showed better of the plant also exhibit activity against Leishmania antiproliferative effects. The main structural difference donovani promastigotes (Mesquita et al. 2005), Trypa- among these six diterpenes is the presence of a casearin nosoma cruzi amastigotes (Espíndola et al. 2004, Mes- C decanoate substituent at C-7 (Fig. 1), which could quita et al. 2005), Plasmodium falciparum strains re- explain its higher cytotoxicity on V-79 cells (Itokawa sistant to chloroquine (Mesquita et al. 2007) and Aedes et al. 1990) likely based on its remarkably larger hydro- aegypti larvae (Rodrigues et al. 2006). Leaf methanolic phobicity and facility to penetrate cytoplasm mem- extract shows anti-hyperlipidemic properties, decreas- brane cell. ing serum triglycerides and lipases (Schoenfelder et al. Recently, two gallic acid-derived compounds iso- 2008) and the water extract displays neuromodulatory lated from C. sylvestris leaves – isobutyl gallate-3,5- effects on central nervous system by the interference on dimethyl ether (IGDE) and methyl gallate-3,5-dimethyl enzymatic action of the purinergic and cholinergic sys- ether (MGDE) – showed significant chemotherapeutic tems since it inhibits Na+-/K+/ATPase and 50-nucleo- potential against Ehrlich and Lewis lung cancer ascite tidase activities (Silva et al. 2006). Additionally, the hy- tumor cells and increased animals’ survival by approx- droalcoholic extract has antinociceptive capability likely imately 90%, though IGDE possesses a slightly su- due to the inhibition of an inflammatory mediator and/or perior activity when compared to MGDE (Da Silva et by the activation of opioid cascade (Mattos et al. 2007). al. 2009). Hydroxybenzoic acids, such as gallic and ellagic acids as well as other polyphenolic molecules CONCLUSION possess cytotoxic activity on human cancer cells (Zhang Several reports have highlighted the medicinal value of et al. 2005, Veluri et al. 2006). In addition, other re- extracts and compounds isolated from Casearia sylves- searches have reported that plants with polyphenolic tris, underscoring a worthy pharmacological arsenal to substances (for instance, flavonoids) protect the cells this plant, bringing evidence to its worldwide popular against injury caused by X-rays, block the progression use and encouraging further searches for novel bioac- of the cell cycle and prostaglandin synthesis, inhibit tive molecules and a better comprehension about its bio- mutations and prevent carcinogenesis in experimental activities. animals (Abdulla and Gruber 2000). Detailed studies revealed that MGDE and IGDE induce higher levels of splenic natural killer (NK) cell ACKNOWLEDGMENTS in tumor-bearing mice (Da Silva et al. 2009). During We wish to thank Conselho Nacional de Desenvol- Erlich ascites tumor growth, the level of NK cell activity vimento Científico e Tecnológico (CNPq), Coordena- in the spleen frequently decreases, which is important ção de Aperfeiçoamento de Pessoal de Nível Superior for tumor proliferation, since NK cells and cytokines (CAPES), Financiadora de Estudos e Projetos (FINEP), (interleukin-2 and interferon-γ ) inhibits the growth of Fundação de Amparo à Pesquisa do Estado de São this murine tumor model (Valadares et al. 2003). More- Paulo (FAPESP) and Banco do Nordeste do Brasil over, evidences show role of interferon-γ in macro- (BNB) for financial support in the form of grants and phage-mediated regulation of host anti-tumor responses fellowship awards.
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