Snake Venom PLA<sub><b>2</b></sub>s Inhibitors Isolated from Brazilian Plants: Synthetic and Natural Molecules

Carvalho, B. M. A.; Santos, J. D. L.; Xavier, B. M.; Almeida, J. R.; Resende, L. M.; Martins, W.; Marcussi, S.; Marangoni, S.; Stábeli, R. G.; Calderon, L. A.; Soares, A. M.; Da Silva, S. L.; Marchi-Salvador, D. P.

doi:https://doi.org/10.1155/2013/153045

BioMed Research International

On this page

Abstract Introduction Acknowledgments References Copyright Related Articles

Special Issue

Animal Toxins and Their Advantages in Biotechnology and Pharmacology

View this Special Issue

Review Article | Open Access

Volume 2013 | Article ID 153045 | https://doi.org/10.1155/2013/153045

Snake Venom PLA₂s Inhibitors Isolated from Brazilian Plants: Synthetic and Natural Molecules

B. M. A. Carvalho,¹J. D. L. Santos,²B. M. Xavier,¹J. R. Almeida,³L. M. Resende,³W. Martins,¹S. Marcussi,⁴S. Marangoni,³R. G. Stábeli,⁵L. A. Calderon,⁵A. M. Soares,⁵and S. L. Da Silva¹ et al.

Academic Editor: Fernando Albericio

Received31 May 2013

Accepted19 Aug 2013

Published22 Sept 2013

Abstract

Ophidian envenomation is an important health problem in Brazil and other South American countries. In folk medicine, especially in developing countries, several vegetal species are employed for the treatment of snakebites in communities that lack prompt access to serum therapy. However, the identification and characterization of the effects of several new plants or their isolated compounds, which are able to inhibit the activities of snake venom, are extremely important and such studies are imperative. Snake venom contains several organic and inorganic compounds; phospholipases A₂ (PLA₂s) are one of the principal toxic components of venom. PLA₂s display a wide variety of pharmacological activities, such as neurotoxicity, myotoxicity, cardiotoxicity, anticoagulant, hemorrhagic, and edema-inducing effects. PLA₂ inhibition is of pharmacological and therapeutic interests as these enzymes are involved in several inflammatory diseases. This review describes the results of several studies of plant extracts and their isolated active principles, when used against crude snake venoms or their toxic fractions. Isolated inhibitors, such as steroids, terpenoids, and phenolic compounds, are able to inhibit PLA₂s from different snake venoms. The design of specific inhibitors of PLA₂s might help in the development of new pharmaceutical drugs, more specific antivenom, or even as alternative approaches for treating snakebites.

1. Introduction

Venomous snakebites represent an important risk for public health worldwide, especially in tropical regions where these accidents are more common. Snake venom is composed by a mixture of inorganic ions (calcium potassium, iron, cobalt, copper, and magnesium), organic compounds like carbohydrate, serotonin, histamine, bradykinin potentiating peptide, disintegrins, and proteins with or without catalytic activity (L-amino acid oxidases, lectins, hyaluronidases, serine proteases, metalloproteases, and phospholipases A₂) [1]. The phospholipase A₂ enzymes (PLA₂s, E.C. 3.1.1.4, and phosphatide sn-2 acylhydrolases) are one of the most important enzymes for its effect. The PLA₂ class includes several polypeptides with similar enzymatic functions; however, these proteins exert a variety of relevant toxic actions, such as neurotoxicity and myotoxicity [2].

Secreted phospholipases A₂ (sPLA₂s) catalyze the hydrolysis of glycerophospholipids in sn-2 position and promote the release of lysophospholipids and fatty acids, such as the arachidonic acid. The arachidonic acid is a precursor of prostaglandins and leukotrienes, and it is involved in inflammatory process characterized by increase by microvascular permeability and oedema formation, leukocyte recruitment into tissues, nociception, and release of inflammatory mediators which mimic a number of systemic and local inflammatory disorders in humans [1–5]. In addition, the excess levels of sPLA₂s were associated with many physiopathological processes as cerebral illnesses, cardiovascular disorders, cancers, asthma, respiratory distress syndrome, and progression of tonsillitis [4–8]. On the other hand, the increased sPLA₂ activity is observed in some brain tumours, in chronic neurological disorders associated with neurodegenerative diseases, such as neural trauma, Alzheimer’s, and Parkinson’s diseases, and may serve as a marker of increases in permeability of the blood-cerebrospinal fluid barrier [9, 10].

PLA₂s show considerable identity in their amino acid sequence [11, 12], but the three-dimensional structure similarity among group II sPLA2s is considerably higher, and this fact shows the importance of the 3D structure for the biological activities [2, 13–16]. Venom of different snake species is used as sources of PLA₂, due to the abundance of these enzymes and the fact that the purification of these molecules is relatively simple [17–19].

The apparent contradiction between structural uniformity and functional diversity, exhibited by PLA₂s, has attracted much interest from the scientific community. According to Ohno and collaborators [20], this diversity of pharmacological and toxic effects may have been evolutionarily acquired by positive Darwinian selection of the coding exons of these activities.

Due to a high degree of structural similarity between the sPLA₂s from snake venom and the human, it is a prerequisite to use the snake venom PLA₂ inhibitors for the design of new drugs for human diseases because the new inhibitory drugs must be related to the transitional state of the enzyme [2, 21]. Small variations among PLA₂ isoforms may be used for the study of structural and functional relationships of these proteins. Moreover, research regarding natural and synthetic inhibitors that are able to neutralize the toxic effects promoted by these enzymes is being carried out in an attempt to explain the physiopathological mechanisms of these molecules [22–24]. Furthermore, knowledge about the mechanism of toxicity exhibited by these proteins may assist the discovery and development of new anti-inflammatory drugs, cellular lesions, and therapies for several diseases, including Parkinson’s, Alzheimer’s, and even cancer [12, 25–29].

Treatment of snakebites is still carried out using traditional antivenom therapy [30]. However, although antivenom therapy is effective for the majority of cases, some side effects exist for these treatments, including adverse reactions on the skin, gastrointestinal tract, and respiratory and circulatory systems [31, 32]. Moreover, snake antivenom therapy is usually unable to prevent the progress of local effects [30]. Given the limitations of traditional therapy, research focusing on the interactions between PLA₂s and their natural or synthetic inhibitors could allow the development of alternative treatments for the toxic and pharmacological effects of snake bites [23, 33]. Plant extracts have become a promising alternative to substitute traditional snake antivenom, which often are unavailable in emergency situations [34, 35]. After studying plants commonly used to treat snakebites in South America, Soares and collaborators [34] reported 56 vegetal species that exhibited anti-inflammatory activity caused by crude snake venom or by their isolated components.

2. PLA₂ Inhibitors Isolated from Plants

Plants are used in traditional medicine to treat the effects of venomous snake bites. Pharmacological studies have shown that fractions of these plant extracts have anti-inflammatory, antiviral, and antivenom properties [36, 37]. The effect of specific molecules from these plant extracts may be attributed to the presence of multiple factors, such as low molecular weight of chemical compounds and the abundance of chemical and pharmacological properties [33].

Borges et al. [38] reported that the aqueous extract of Casearia sylvestris (Flacourtiaceae), a native vegetal species found in Brazilian open pastures, had the ability to inhibit myotoxic, anticoagulant, and edema-inducing activities from Bothrops moojeni, B. pirajai, B. neuwiedi, and B. jararacussu venom and its Asp49 and Lys49-PLA₂ isolated toxins. In addition, Borges and collaborators [39] emphasized that C. sylvestris was able to neutralize hemorrhagic activity caused by the B. pirajai, B. jararacussu, B. asper, B. moojeni, and B. neuwiedi venom. Cavalcante and collaborators [22] showed that the C. sylvestris aqueous extract demonstrated protective effects against muscle damage induced by two Lys49-PLA₂ toxins (PrTX-I from B. pirajai and BthTX-I from B. jararacussu snake venom) and prevented the neuromuscular blockage induced by all PLA₂ toxins.

Mandevilla velutina (Apocynaceae) is a perennial plant from the Brazilian cerrado that has been studied for its anti-inflammatory activity, as well as its antagonist effect on bradykinin, a vasodilator [40]. These authors reported that the aqueous extract of this plant was an effective inhibitor of phospholipase A₂ activity and some toxic effects, such as hemorrhage, caused by venom from snakes of the Bothrops and Crotalus genus. In a posterior study, the same research group reported that extracts from Mandevilla illustris were able to completely inhibit the activity of the Crotoxin B, the basic Asp49-PLA₂, isolated from Crotalus durissus terrificus venom [41].

The antihemorrhagic properties of the aqueous extract of Pentaclethra macroloba (Fabaceae), an ethnomedicinal plant found in the Amazon region, were evaluated against snake venom and displayed a full inhibition of hemorrhagic and nucleolytic activities induced by several snake venom. Additionally, a partial inhibition of myotoxic, lethal, enzymatic and edema activities of snake venom, and their isolated PLA₂s was observed [42].

Almeida and collaborators [43] showed that the aqueous extract of Tabernaemontana catharinensis (Apocynaceae), which is encountered in some countries of South America, was able to inhibit the crotoxin complex, isolated from C. d. terrificus venom, and was also able to partially neutralize the myotoxicity of B. jararacussu snake venom and its basic PLA₂s [44].

The aqueous extract of the aerial parts of Bauhinia forficate (Fabaceae), a species native to Asia and well adapted and developed in several regions of Brazil, was assayed against the fibrinogenolytic and coagulant activities of C. d. terrificus and B. jararacussu crude venom and was found to neutralize these effects. Moreover, the extract efficiently inhibited the edema induced by C. d. terrificus venom and its isolated PLA₂ [45].

Mendes and collaborators [46] reported that the aqueous extract of Schizolobium parahyba (Fabaceae), a plant found in the Mata Atlântica of southeastern Brazil, contains compounds that can inhibit some enzymatic and biological activities induced by Bothrops pauloensis (current Bothropoides paulensis) and C. d. terrificus snake venom as well as by their isolated neuwiedase toxins (metalloproteinase), BnSP-7 (basic Lys49-PLA₂ from B. paulensis venom), and Crotoxin B.

The ethanolic extract of the aerial parts of Blutaparon portulacoides (Amaranthaceae), an herbaceous plant that occurs mainly in the Atlantic bush, caused a reduction in edema formation and in the leukocyte influx induced by Lys49-PLA₂ and isolated from B. jararacussu venom [47].

In 2005, Maiorano and collaborators [48] evaluated aqueous extracts prepared from dried or fresh roots, stems, or leaves of Mikania glomerata (Asteraceae), a plant found in the Mata Atlântica in Southeastern Brazil and popularly known as “Guaco.” The M. glomerata extract efficiently neutralized different toxic, pharmacological, and enzymatic effects induced by Bothrops and Crotalus snake venom. The phospholipase A₂ activity and the edema induced by C. d. terrificus venom were inhibited by approximately 100 and 40%, respectively, and this inhibition was also partially observed with the Bothrops venom. The hemorrhagic activities of B. alternatus, B. moojeni, B. neuwiedi, and B. jararacussu venom were significantly inhibited by M. glomerata extract, while the clotting activities of C. d. terrificus, B. jararacussu, and B. neuwiedi venom were totally inhibited. In addition, Floriano and collaborators [49] reported clinical and laboratory alterations in mice caused by the combination of M. glomerata leaf extract and antiophidian serum against C. d. terrificus venom.

Nazato and collaborators [50] found that the methanolic extract of the bark from Dipteryx alata (Fabaceae) (a native species of the Brazilian Savanna, found principally in Minas Gerais, Goiás, Federal District, and Mato Grosso) decreased the neurotoxicity and myotoxicity of B. jararacussu crude venom. In another study by Puebla and collaborators [51], the extract from D. alata was fractionated and its compounds were evaluated against the neuromuscular blockade caused by B. jararacussu venom.

The ability of the ethanolic extract of Hypericum brasiliense (Hypericaceae), a plant found mainly in the southeastern and southern regions of Brazil, to neutralize some effects induced by B. jararaca venom was investigated using biological assays. H. brasiliense extracts were able to inhibit some pharmacological effects such as lethality, edema, hemorrhage, hemolysis and, proteolysis, as well as fibrinogen or plasma clotting [52].

In 2012, Dey and De [53] published a review that evaluated several pharmacological studies on plant efficacies against snakebites. The authors compiled studies from a number of plants or their fractionsthat were active against snake venom and concluded that folk knowledge is relevant. However, clinical tests should be performed with these plant extracts or fractions to assess the effect of the compounds used for the treatment of snakebites.

Recently, Samy et al. [54] published an extensive revision on the therapeutic application of natural inhibitors of snake venom PLA₂s, covering molecules from the primary metabolism of different organisms, such as glycoproteins (PLIs), peptides, and lipids, as well as from secondary metabolism, exemplified by terpenoids, alkaloids, flavonoids, and other molecules. These authors concluded that the biotechnological potential of PLA₂ inhibitors may provide therapeutic molecular models with antiophidian activity to supplement conventional serum therapy or for the development of novel antivenom therapeutics. Additionally, inhibitors isolated from medicinal plants may also be an essential tool in isolated communities [23, 54].

3. Structural Characterization of PLA₂ Inhibitors

The main classes of PLA₂ inhibitors are the phenolic compounds, which include flavonoids, coumestans and alkaloids, steroids and terpenoids (mono-, di-, and triterpenes), and polyphenols (vegetable tannins). There is also mention in the literature of other molecules such as carbohydrates, lipids, and proteins, although this paper emphasizes molecules originating from plant secondary metabolism.

3.1. Phenolic Compounds

3.1.1. Flavonoids

Polyphenolic secondary metabolites are commonly able to bind to biological polymers, and some of these have been shown to inhibit PLA₂s. Examples include quercetin, a strong lipoxygenase inhibitor, naringenin, artemetin, kaempferol, and galangin, among several other flavonoids. Primetin (Figure 1), 5,8-dihydroxyflavone, isolated from Primula sp. (Primulaceae), is known for its ability to inhibit toxins from snake venom; its structural form may be seen in Figure 1. Flavonoids usually exert their inhibitory effect via hydrophobic interactions with the A and B rings and aromatic or hydrophobic amino acid residues in the protein [36, 55, 56].

3.1.2. Coumestans

Eclipta alba (Asteraceae) is a native plant from Brazil and other tropical and subtropical areas of the world whose medicinal properties are widely known. E. alba was genetically engineered using Agrobacterium rhizogenes LB9402 to enhance the production of secondary wedelolactone metabolites, which are coumestan compounds with activity against basic PLA₂s. This mutant strain was found to reduce the phospholipase A₂ activities and myotoxic and neurotoxic effects of the C. d. terrificus and B. jararacussu snake venom [37, 57]. Analogs of wedelolactone molecule (Figure 1) were able to antagonize the release of creatine kinase induced by B. jararacussu venom even at concentrations as low as 30 μM [58, 59].

3.1.3. Alkaloids

Batina and collaborators [60] isolated an alkaloid from Tabernaemontana catharinensis (Apocynaceae) named 12-methoxy-4-methylvoachalotine (Figure 1) and reported a strong inhibitory effect against lethality and myotoxic activities induced by C. d. terrificus venom.

3.1.4. Other Phenolic Compounds

Ar-Turmerone (Figure 1) is a phenolic compound isolated from the Curcuma longa (Zingiberaceae) plant that has a strong effect against the hemorrhage and lethality caused by B. jararaca and C. d. terrificus snake venom [61].

Extracts from Piper umbellatum and P. peltatum (Piperaceae)were shown to inhibit the myotoxic activities of PLA₂s isolated from Bothrops snake venom [62]. Fractionation of these plant extracts revealed that 4-nerolidylcatechol, a hydroxylated phenolic compound (Figure 1), was responsible for at least part of the inhibitory effect against groups I, II, and III of PLA₂s.

In 2008, Da Silva and collaborators [63] studied the half maximal inhibitory concentration (IC₅₀) of ellagic acid (Figure 1), extracted from C. sylvestris, against BthTX-II, a basic Asp49-PLA₂ from B. jararacussu snake venom, and concluded that this compound was effective at competitively inhibiting the induction of edema, myotoxicity, and enzymatic activities, incurred by this PLA₂.

The first structural analysis of aristolochic acid (Figure 1), isolated from Aristolochia sp. (Aristolochiaceae), was performed by Vishwanath and Gowda [64]. In this study, the interaction of aristolochic acid, an alkaloid, with PLA₂ from Vipera russelli was characterized as noncompetitive inhibitive. This compound has also been shown to reduce the induction of edema by this enzyme. Additionally, Vishwanath et al. [65] emphasized that the interaction between aristolochic acid, from Aristolochia radix, and three PLA₂s from Trimeresurus flavoviridis resulted in the inhibition of hemolytic and edema induction by competitive inhibition. Chandra and collaborators [66] reported the crystal structure of the complex formed between the PLA₂ isolated from V. russelli venom and aristolochic acid. In this study, the interaction between aristolochic acid and PLA₂ was competitive, and the final model consists of a dimer of PLA₂ and one molecule of aristolochic acid located in the binding site of molecule A; this interaction was stabilized by three hydrogen bonds and hydrophobic contacts.

Chlorogenic and caffeic acids (Figure 1) can interact with proteins via hydrophobic contacts and hydrogen bonds, inhibiting enzyme function and acting as antidotes. Strong interactions may induce conformational changes in the protein structure [26]. In 2011, Shimabuku and collaborators [67] crystallized PrTX-I (basic Lys49-PLA₂ from B. pirajai snake venom) in the presence of the inhibitor, caffeic acid, and the electron-density map which unambiguously indicated the presence of three caffeic acid molecules interacting with the C-terminus of the protein.

Rosmarinic acid (Figure 1) is a hydroxylated phenolic compound isolated from Cordia verbenacea (Boraginaceae). This compound demonstrates antimyotoxic properties and inhibits edema induced by crude B. jararacussu snake venom and its basic PLA₂s [36, 55]. The three-dimensional structure of the PrTX-I, rosmarinic acid complex, was elucidated by Santos and collaborators [68], where rosmarinic acid was observed located at the entrance of the hydrophobic channel monomer A of the PrTX-I dimer via an interaction between hydrogen bonds and hydrophobic contacts in the same monomer. Interactions were also observed between rosmarinic acid and a residue of the C-terminal region of the monomer B. The interaction between the rosmarinic acid molecule with the hydrophobic channel (monomer A) and the C-terminal region (myotoxic site, monomer B) suggests two mechanisms of myotoxicity inhibition [68].

3.2. Steroid Compounds

Sterol and cholesterol molecules present well-known antidote activities against snake venom. Steroids can form complexes that are stabilized via Van der Waals interactions, as well as by hydrophobic interactions [37]. Antimyotoxic and antihemorrhagic effects of the Eclipta prostrata (Asteraceae) extract and its components, sitosterol and stigmasterol (Figure 1), were observed against B. jararaca, B. jararacussu, and Lachesis muta snake venom [37, 69]. Previously, Mors [70] reported that sitosterol and stigmasterol, isolated from E. prostrata, prevented the lethality of the C. d. terrificus venom in a dose-dependent manner.

3.3. Terpenoids

The neoclerodane, diterpenoid, isolated from the aerial parts of Baccharis trimera (Asteraceae), demonstrate anti-hemorrhagic and antiprotolithic properties against Bothrops snake venom [71]

Several pentacyclic triterpenes, such as oleanolic acid, lupeol, ursolic acid, taraxerol, taraxasterol, α,β-amyrin, and friedeline, exhibit activity against snake venom [37]. Triterpenoids, isolated from Betula alba (Betulaceae), including pentacyclic triterpenes betulin and betulinic acid (Figure 1), exhibited antiphospholipasic A₂ activity. Docking (in silico experiments) indicated betulinic acid as the best PLA₂ inhibitor, due to its direct insertion in the catalytic site on the enzyme, with a very low energy value [55].

3.4. Synthetic Inhibitors

Edunol (Figure 1) is a pterocarpan with a chemical structure similar to those of the inhibitors extracted from the roots of Harpalyce brasiliana (Fabaceae). Edunol was obtained via chemical synthesis, and the compound showed anti-myotoxic, anti-proteolytic, and anti-PLA₂ activities against B. jararacussu crude venom [55, 72].

Elaidoylamide, the amide of trans-9-octadecenoic acid (Figure 1), is a powerful synthetic inhibitor of a neurotoxic Asp49-PLA₂ from Vipera ammodytes meridionalis venom. In 2003, Georgieva and collaborators [73] isolated the neurotoxic complex from V. a. meridionalis venom, dissociated the basic PLA₂ from the complex, and crystallized it with elaidoylamide. This final structure contained two identical homodimers and one molecule of elaidoylamide bound simultaneously to the substrate-binding sites of each homodimer [74].

Villar and collaborators [33] demonstrated that synthetic inhibitor derivatives from nitrostyrene that contain typical nitro groups at the ortho-, meta-, and para- positions on the aromatic ring were more efficient against the enzymatic, edematogenic, and myotoxic activities of PLA₂s from B. jararacussu venom. Da Silva and collaborators [75, 76], performing molecular modeling studies between Asp49-PLA₂ from C. adamanteus venom and synthetic derivatives polyhydroxy phenolic compounds, concluded that some conformations of these groups might positively influence enzymatic activity inhibition.

Isolated inhibitors (natural or synthetic) can be important tools for understanding the mechanisms of action of PLA₂s from snake venom, and, consequently, these results might be helpful for the design of a drug that specifically inhibits PLA₂s. However, the synthesis of compounds analogous to their natural equivalents, based on chemical characteristics or with minor structural modifications, is often necessary. The synthesis of compounds could be justified by the low amount of these compounds available in vegetal extracts or to adjust some specific chemical characteristics. For this reason, some researchers have isolated and characterized new compounds or produced synthetic analogues for use in the commercial production of pharmaceutical drugs.

Acknowledgments

The authors gratefully acknowledge financial support from Coordenação de Aperfeiçoamento de Nível Superior (CAPES), Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

References

D. C. I. Koh, A. Armugam, and K. Jeyaseelan, “Snake venom components and their applications in biomedicine,” Cellular and Molecular Life Sciences, vol. 63, no. 24, pp. 3030–3041, 2006.
View at: Publisher Site | Google Scholar
R. K. Arni and R. J. Ward, “Phospholipase A₂—a structural review,” Toxicon, vol. 34, no. 8, pp. 827–841, 1996.
View at: Publisher Site | Google Scholar
C. D. Funk, “Prostaglandins and Leukotrienes: advances in eicosanoid biology,” Science, vol. 294, no. 5548, pp. 1871–1875, 2001.
View at: Publisher Site | Google Scholar
F. S. Markland Jr, “Snake venoms and the hemostatic system,” Toxicon, vol. 36, no. 12, pp. 1749–1800, 1998.
View at: Publisher Site | Google Scholar
S. L. da Silva, M. Comar Jr., K. M. T. Oliveira et al., “Molecular modeling of the inhibition of enzyme PLA₂ from snake venom by dipyrone and 1-phenyl-3-methyl-5-pyrazolone,” International Journal of Quantum Chemistry, vol. 108, no. 13, pp. 2576–2585, 2008.
View at: Publisher Site | Google Scholar
Z. Mallat, G. Lambeau, and A. Tedgui, “Lipoprotein-associated and secreted phospholipases A₂ in cardiovascular disease: roles as biological effectors and biomarkers,” Circulation, vol. 122, no. 21, pp. 2183–2200, 2010.
View at: Publisher Site | Google Scholar
D. de Luca, A. Minucci, P. Cogo et al., “Secretory phospholipase A₂ pathway during pediatric acute respiratory distress syndrome: a preliminary study,” Pediatric Critical Care Medicine, vol. 12, no. 1, pp. e20–e24, 2011.
View at: Publisher Site | Google Scholar
R. Ezzeddini, M. Darabi, B. Ghasemi et al., “Circulating phospholipase A₂ activity in obstructive sleep apnea and recurrent tonsillitis,” International Journal of Pediatric Otorhinolaryngology, vol. 76, no. 4, pp. 471–474, 2012.
View at: Publisher Site | Google Scholar
S. Chalbot, H. Zetterberg, K. Blennow et al., “Blood-cerebrospinal fluid barrier permeability in Alzheimer's disease,” Journal of Alzheimer's Disease, vol. 25, no. 3, pp. 505–515, 2011.
View at: Publisher Site | Google Scholar
A. A. Farooqui, M. L. Litsky, T. Farooqui, and L. A. Horrocks, “Inhibitors of intracellular phospholipase A₂ activity: their neurochemical effects and therapeutical importance for neurological disorders,” Brain Research Bulletin, vol. 49, no. 3, pp. 139–153, 1999.
View at: Publisher Site | Google Scholar
S. S. Teixeira, L. B. Silveira, F. M. N. da Silva et al., “Molecular characterization of an acidic phospholipase A₂ from Bothrops pirajai snake venom: synthetic C-terminal peptide identifies its antiplatelet region,” Archives of Toxicology, vol. 85, no. 10, pp. 1219–1233, 2011.
View at: Publisher Site | Google Scholar
L. Romero, S. Marcussi, D. P. Marchi-Salvador et al., “Enzymatic and structural characterization of a basic phospholipase A₂ from the sea anemone Condylactis gigantea,” Biochimie, vol. 92, no. 8, pp. 1063–1071, 2010.
View at: Publisher Site | Google Scholar
L. C. Corrêa, D. P. Marchi-Salvador, A. C. O. Cintra, S. V. Sampaio, A. M. Soares, and M. R. M. Fontes, “Crystal structure of a myotoxic Asp49-phospholipase A₂ with low catalytic activity: insights into Ca²⁺ independent catalytic mechanism,” Biochimica et Biophysica Acta, vol. 1784, no. 4, pp. 591–599, 2008.
View at: Publisher Site | Google Scholar
D. P. Marchi-Salvador, L. C. Corrêa, A. J. Magro, C. Z. Oliveira, A. M. Soares, and M. R. M. Fontes, “Insights into the role of oligomeric state on the biological activities of crotoxin: crystal structure of a tetrameric phospholipase A₂ formed by two isoforms of crotoxin B from Crotalus durissus terrificus venom,” Proteins, vol. 72, no. 3, pp. 883–891, 2008.
View at: Publisher Site | Google Scholar
D. P. Marchi-Salvador, C. A. H. Fernandes, L. B. Silveira, A. M. Soares, and M. R. M. Fontes, “Crystal structure of a phospholipase A₂ homolog complexed with p-bromophenacyl bromide reveals important structural changes associated with the inhibition of myotoxic activity,” Biochimica et Biophysica Acta, vol. 1794, no. 11, pp. 1583–1590, 2009.
View at: Publisher Site | Google Scholar
C. A. H. Fernandes, D. P. Marchi-Salvador, G. H. M. Salvador et al., “Comparison between apo and complexed structures of bothropstoxin-I reveals the role of Lys122 and Ca²⁺-binding loop region for the catalytically inactive Lys49-PLA₂s,” Journal of Structural Biology, vol. 171, no. 1, pp. 31–43, 2010.
View at: Publisher Site | Google Scholar
J. L. C. Cardoso, F. O. S. França, F. H. Wen, C. M. S. Málaque, and V. Haddad Jr, Animais peçonhentos no Brasil: biologia clinica e terapêutica dos acidentes, Sarvier, São Paulo, Brazil, 2003.
D. C. S. Damico, M. A. C. Höfling, M. Cintra et al., “Pharmacological study of edema and myonecrosis in mice induced by venom of the bushmaster snake (Lachesis muta muta) and its basic Asp49 phospholipase A₂ (LmTX-I),” The Protein Journal, vol. 27, no. 6, pp. 384–391, 2008.
View at: Publisher Site | Google Scholar
D. C. S. Damico, T. Vassequi-Silva, F. D. Torres-Huaco et al., “LmrTX, a basic PLA₂ (D49) purified from Lachesis muta rhombeata snake venom with enzymatic-related antithrombotic and anticoagulant activity,” Toxicon, vol. 60, pp. 773–781, 2012.
View at: Google Scholar
M. Ohno, R. Ménez, T. Ogawa et al., “Molecular evolution of snake toxins: is the functional diversity of snake toxins associated with a mechanism of accelerated evolution?” Progress in Nucleic Acid Research and Molecular Biology, vol. 59, pp. 307–364, 1998.
View at: Google Scholar
E. Valentin and G. Lambeau, “What can venom phospholipases A₂ tell us about the functional diversity of mammalian secreted phospholipases A₂?” Biochimie, vol. 82, no. 9-10, pp. 815–831, 2000.
View at: Publisher Site | Google Scholar
W. L. G. Cavalcante, T. O. Campos, M. dal Pai-Silva et al., “Neutralization of snake venom phospholipase A₂ toxins by aqueous extract of Casearia sylvestris (Flacourtiaceae) in mouse neuromuscular preparation,” Journal of Ethnopharmacology, vol. 112, no. 3, pp. 490–497, 2007.
View at: Publisher Site | Google Scholar
S. Marcussi, C. D. Sant'Ana, C. Z. Oliveira et al., “Snake venom phospholipase A₂ inhibitors: medicinal chemistry and therapeutic potential,” Current Topics in Medicinal Chemistry, vol. 7, no. 8, pp. 743–756, 2007.
View at: Publisher Site | Google Scholar
C. Z. Oliveira, D. L. Menaldo, S. Marcussi et al., “An α-type phospholipase A₂ inhibitor from Bothrops jararacussu snake plasma: structural and functional characterization,” Biochimie, vol. 90, no. 10, pp. 1506–1514, 2008.
View at: Publisher Site | Google Scholar
B. S. Cummings, J. McHowat, and R. G. Schnellmann, “Phospholipase A₂s in cell injury and death,” The Journal of Pharmacology and Experimental Therapeutics, vol. 294, no. 3, pp. 793–799, 2000.
View at: Google Scholar
C. Araya and B. Lomonte, “Antitumor effects of cationic synthetic peptides derived from Lys49 phospholipase A₂ homologues of snake venoms,” Cell Biology International, vol. 31, no. 3, pp. 263–268, 2007.
View at: Publisher Site | Google Scholar
B. Lomonte, Y. Angulo, and E. Moreno, “Synthetic peptides derived from the C-Terminal region of Lys49 phospholipase A₂ homologues from viperidae snake venoms: biomimetic activities and potential applications,” Current Pharmaceutical Design, vol. 16, no. 28, pp. 3224–3230, 2010.
View at: Publisher Site | Google Scholar
R. Perumal Samy and P. Gopalakrishnakone, “Therapeutic potential of plants as anti-microbials for drug discovery,” Evidence-Based Complementary and Alternative Medicine, vol. 7, no. 3, pp. 283–294, 2010.
View at: Publisher Site | Google Scholar
C. Z. Oliveira, N. A. Santos-Filho, D. L. Menaldo et al., “Structural and functional characterization of a γ-type phospholipase A₂ inhibitor from Bothrops jararacussu Snake Plasma,” Current Topics in Medicinal Chemistry, vol. 11, no. 20, pp. 2509–2519, 2011.
View at: Google Scholar
J. M. Gutiérrez, B. Lomonte, G. León, A. Rucavado, F. Chaves, and Y. Angulo, “Trends in snakebite envenomation therapy: scientific, technological and public health considerations,” Current Pharmaceutical Design, vol. 13, no. 28, pp. 2935–2950, 2007.
View at: Publisher Site | Google Scholar
M. García, M. Monge, G. León et al., “Effect of preservatives on IgG aggregation, complement-activating effect and hypotensive activity of horse polyvalent antivenom used in snakebite envenomation,” Biologicals, vol. 30, no. 2, pp. 143–151, 2002.
View at: Publisher Site | Google Scholar
J. M. Schmidt, “Antivenom therapy for snakebites in children: is there evidence?” Current Opinion in Pediatrics, vol. 17, no. 2, pp. 234–238, 2005.
View at: Publisher Site | Google Scholar
J. A. F. P. Villar, F. T. D. Lima, C. L. Veber et al., “Synthesis and evaluation of nitrostyrene derivative compounds, new snake venom phospholipase A₂ inhibitors,” Toxicon, vol. 51, no. 8, pp. 1467–1478, 2008.
View at: Publisher Site | Google Scholar
A. M. Soares, A. H. Januário, M. V. Lourenço, A. M. S. Pereira, and P. S. Pereira, “Neutralizing effects of Brazilian plants against snake venoms,” Drugs of the Future, vol. 29, no. 11, pp. 1105–1117, 2004.
View at: Publisher Site | Google Scholar
M. L. da Silva, S. Marcussi, R. S. Fernandes et al., “Anti-snake venom activities of extracts and fractions from callus cultures of Sapindus saponaria,” Pharmaceutical Biology, vol. 50, no. 3, pp. 366–375, 2012.
View at: Publisher Site | Google Scholar
F. K. Ticli, L. I. S. Hage, R. S. Cambraia et al., “Rosmarinic acid, a new snake venom phospholipase A₂ inhibitor from Cordia verbenacea (Boraginaceae): antiserum action potentiation and molecular interaction,” Toxicon, vol. 46, no. 3, pp. 318–327, 2005.
View at: Publisher Site | Google Scholar
W. B. Mors, M. C. Nascimento, B. M. R. Pereira, and N. A. Pereira, “Plant natural products active against snake bite—the molecular approach,” Phytochemistry, vol. 55, no. 6, pp. 627–642, 2000.
View at: Publisher Site | Google Scholar
M. H. Borges, A. M. Soares, V. M. Rodrigues et al., “Effects of aqueous extract of Casearia sylvestris (Flacourtiaceae) on actions of snake and bee venoms and on activity of phospholipases A₂,” Comparative Biochemistry and Physiology B, vol. 127, no. 1, pp. 21–30, 2000.
View at: Publisher Site | Google Scholar
M. H. Borges, A. M. Soares, V. M. Rodrigues et al., “Neutralization of proteases from Bothrops snake venoms by the aqueous extract from Casearia sylvestris (Flacourtiaceae),” Toxicon, vol. 39, no. 12, pp. 1863–1869, 2001.
View at: Publisher Site | Google Scholar
R. Biondo, A. M. S. Pereira, S. Marcussi, P. S. Pereira, S. C. França, and A. M. Soares, “Inhibition of enzymatic and pharmacological activities of some snake venoms and toxins by Mandevilla velutina (Apocynaceae) aqueous extract,” Biochimie, vol. 85, no. 10, pp. 1017–1025, 2003.
View at: Publisher Site | Google Scholar
R. Biondo, A. M. Soares, B. W. Bertoni, S. C. França, and A. M. S. Pereira, “Direct organogenesis of Mandevilla illustris (Vell) Woodson and effects of its aqueous extract on the enzymatic and toxic activities of Crotalus durissus terrificus snake venom,” Plant Cell Reports, vol. 22, no. 8, pp. 549–552, 2004.
View at: Publisher Site | Google Scholar
J. O. da Silva, J. S. Coppede, V. C. Fernandes et al., “Antihemorrhagic, antinucleolytic and other antiophidian properties of the aqueous extract from Pentaclethra macroloba,” Journal of Ethnopharmacology, vol. 100, no. 1-2, pp. 145–152, 2005.
View at: Publisher Site | Google Scholar
L. Almeida, A. C. O. Cintra, E. L. G. Veronese et al., “Anticrotalic and antitumoral activities of gel filtration fractions of aqueous extract from Tabernaemontana catharinensis (Apocynaceae),” Comparative Biochemistry and Physiology C, vol. 137, no. 1, pp. 19–27, 2004.
View at: Publisher Site | Google Scholar
E. L. G. Veronese, L. E. Esmeraldino, A. P. F. Trombone et al., “Inhibition of the myotoxic activity of Bothrops jararacussu venom and its two major myotoxins, BthTX-I and BthTX-II, by the aqueous extract of Tabernaemontana catharinensis A. DC. (Apocynaceae),” Phytomedicine, vol. 12, no. 1-2, pp. 123–130, 2005.
View at: Publisher Site | Google Scholar
C. Z. Oliveira, V. A. Maiorano, S. Marcussi et al., “Anticoagulant and antifibrinogenolytic properties of the aqueous extract from Bauhinia forficata against snake venoms,” Journal of Ethnopharmacology, vol. 98, no. 1-2, pp. 213–216, 2005.
View at: Publisher Site | Google Scholar
M. M. Mendes, C. F. Oliveira, D. S. Lopes et al., “Anti-snake venom properties of Schizolobium parahyba (Caesalpinoideae) aqueous leaves extract,” Phytotherapy Research, vol. 22, no. 7, pp. 859–866, 2008.
View at: Publisher Site | Google Scholar
I. C. Pereira, A. M. Barbosa, M. J. Salvador et al., “Anti-inflammatory activity of Blutaparon portulacoides ethanolic extract against the inflammatory reaction induced by Bothrops jararacussu venom and isolated myotoxins BthTX-I and II,” Journal of Venomous Animals and Toxins Including Tropical Diseases, vol. 15, no. 3, pp. 527–545, 2009.
View at: Publisher Site | Google Scholar
V. A. Maiorano, S. Marcussi, M. A. F. Daher et al., “Antiophidian properties of the aqueous extract of Mikania glomerata,” Journal of Ethnopharmacology, vol. 102, no. 3, pp. 364–370, 2005.
View at: Publisher Site | Google Scholar
R. S. Floriano, R. M. B. Nogueira, M. Sakate et al., “Effect of Mikania glomerata (Asteraceae) leaf extract combined with anti-venom serum on experimental Crotalus durissus (Squamata: Viperidae) envenomation in rats,” Revista de Biología Tropical, vol. 57, no. 4, pp. 929–937, 2009.
View at: Google Scholar
V. S. Nazato, L. Rubem-Mauro, N. A. G. Vieira et al., “In vitro antiophidian properties of Dipteryx alataVogel bark extracts,” Molecules, vol. 15, no. 9, pp. 5956–5970, 2010.
View at: Publisher Site | Google Scholar
P. Puebla, Y. Oshima-Franco, L. M. Franco et al., “Chemical constituents of the bark of Dipteryx alata vogel, an active species against Bothrops jararacussu venom,” Molecules, vol. 15, no. 11, pp. 8193–8204, 2010.
View at: Publisher Site | Google Scholar
M. Assafim, E. C. Coriolano, S. E. Benedito et al., “Hypericum brasiliense plant extract neutralizes some biological effects of Bothrops jararaca snake venom,” Journal of Venom Research, vol. 2, pp. 11–16, 2011.
View at: Google Scholar
A. Dey and J. N. De, “Phytopharmacology of antiophidian botanicals: a review,” International Journal of Pharmacology, vol. 8, no. 2, pp. 62–79, 2012.
View at: Publisher Site | Google Scholar
R. P. Samy, P. Gopalakrishnakone, and V. T. Chow, “Therapeutic application of natural inhibitors against snake venom phospholipase A₂,” Bioinformation, vol. 8, pp. 48–57, 2012.
View at: Google Scholar
A. M. Soares, F. K. Ticli, S. Marcussi et al., “Medicinal plants with inhibitory properties against snake venoms,” Current Medicinal Chemistry, vol. 12, no. 22, pp. 2625–2641, 2005.
View at: Publisher Site | Google Scholar
J. Lättig, M. Böhl, P. Fischer et al., “Mechanism of inhibition of human secretory phospholipase A₂ by flavonoids: rationale for lead design,” Journal of Computer-Aided Molecular Design, vol. 21, no. 8, pp. 473–483, 2007.
View at: Publisher Site | Google Scholar
L. C. Diogo, R. S. Fernandes, S. Marcussi et al., “Inhibition of snake venoms and phospholipases A₂ by extracts from native and genetically modified Eclipta alba: isolation of active coumestans,” Basic & Clinical Pharmacology & Toxicology, vol. 104, no. 4, pp. 293–299, 2009.
View at: Publisher Site | Google Scholar
A. J. M. da Silva, P. A. Melo, N. M. V. Silva et al., “Synthesis and preliminary pharmacological evaluation of coumestans with different patterns of oxygenation,” Bioorganic & Medicinal Chemistry Letters, vol. 11, no. 3, pp. 283–286, 2001.
View at: Publisher Site | Google Scholar
R. C. S. Nunomura, V. G. Oliveira, S. L. da Silva, and S. M. Nunomura, “Characterization of bergenin in Endopleura uchi bark and its anti-inflammatory activity,” Journal of the Brazilian Chemical Society, vol. 20, no. 6, pp. 1060–1064, 2009.
View at: Google Scholar
M. F. Batina, A. C. O. Cintra, E. L. G. Veronese et al., “Inhibition of the lethal and myotoxic activities of Crotalus durissus terrificus venom by Tabernaemontana catharinensis: identification of one of the active components,” Planta Medica, vol. 66, no. 5, pp. 424–428, 2000.
View at: Publisher Site | Google Scholar
L. A. F. Ferreira, O. B. Henriques, A. A. S. Andreoni et al., “Antivenom and biological effects of ar-turmerone isolated from Curcuma longa (Zingiberaceae),” Toxicon, vol. 30, no. 10, pp. 1211–1218, 1992.
View at: Publisher Site | Google Scholar
V. Núñez, V. Castro, R. Murillo, L. A. Ponce-Soto, I. Merfort, and B. Lomonte, “Inhibitory effects of Piper umbellatum and Piper peltatum extracts towards myotoxic phospholipases A₂ from Bothrops snake venoms: isolation of 4-nerolidylcatechol as active principle,” Phytochemistry, vol. 66, no. 9, pp. 1017–1025, 2005.
View at: Publisher Site | Google Scholar
S. L. da Silva, A. K. Calgarotto, J. S. Chaar, and S. Marangoni, “Isolation and characterization of ellagic acid derivatives isolated from Casearia sylvestris SW aqueous extract with anti-PLA₂ activity,” Toxicon, vol. 52, no. 6, pp. 655–666, 2008.
View at: Publisher Site | Google Scholar
B. S. Vishwanath and T. V. Gowda, “Interaction of aristolochic acid with Vipera russelli phospholipase A₂: its effect on enzymatic and pathological activities,” Toxicon, vol. 25, no. 9, pp. 929–937, 1987.
View at: Google Scholar
B. S. Vishwanath, R. M. Kini, and T. V. Gowda, “Characterization of three edema-inducing phospholipase A₂ enzymes from habu (Trimeresurus flavoviridis) venom and their interaction with the alkaloid aristolochic acid,” Toxicon, vol. 25, no. 5, pp. 501–515, 1987.
View at: Google Scholar
V. Chandra, J. Jasti, P. Kaur, A. Srinivasan, C. H. Betzel, and T. P. Singh, “Structural basis of phospholipase A₂ inhibition for the synthesis of prostaglandins by the plant alkaloid aristolochic acid from a 1.7 Å crystal structure,” Biochemistry, vol. 41, no. 36, pp. 10914–10919, 2002.
View at: Publisher Site | Google Scholar
P. S. Shimabuku, C. A. H. Fernandes, A. J. Magro, T. R. Costa, A. M. Soares, and M. R. M. Fontes, “Crystallization and preliminary X-ray diffraction analysis of a Lys49-phospholipase A₂ complexed with caffeic acid, a molecule with inhibitory properties against snake venoms,” Acta Crystallographica F, vol. 67, no. 2, pp. 249–252, 2011.
View at: Publisher Site | Google Scholar
J. I. Santos, F. F. Cardoso, A. M. Soares, M. dal Pai Silva, M. Gallacci, and M. R. M. Fontes, “Structural and functional studies of a bothropic myotoxin complexed to rosmarinic acid: new insights into Lys49-PLA₂ inhibition,” PLoS One, vol. 6, no. 12, Article ID e28521, pp. 1–11, 2011.
View at: Publisher Site | Google Scholar
A. D. L. Souza, E. Rodrigues-Filho, A. Q. L. Souza et al., “Koninginins, phospholipase A₂ inhibitors from endophytic fungus Trichoderma koningii,” Toxicon, vol. 51, no. 2, pp. 240–250, 2008.
View at: Publisher Site | Google Scholar
W. B. Mors, M. C. do Nascimento, J. P. Parente, M. H. da Silva, P. A. Melo, and G. Suarez-Kurtz, “Neutralization of lethal and myotoxic activities of South American rattlesnake venom by extracts and constituents of the plant Eclipta prostrata (Asteraceae),” Toxicon, vol. 27, no. 9, pp. 1003–1009, 1989.
View at: Google Scholar
A. H. Januário, S. L. Santos, S. Marcussi et al., “Neo-clerodane diterpenoid, a new metalloprotease snake venom inhibitor from Baccharis trimera (Asteraceae): anti-proteolytic and anti-hemorrhagic properties,” Chemico-Biological Interactions, vol. 150, no. 3, pp. 243–251, 2004.
View at: Publisher Site | Google Scholar
A. J. M. da Silva, A. L. Coelho, A. B. C. Simas et al., “Synthesis and pharmacological evaluation of prenylated and benzylated pterocarpans against snake venom,” Bioorganic & Medicinal Chemistry Letters, vol. 14, no. 2, pp. 431–435, 2004.
View at: Publisher Site | Google Scholar
D. N. Georgieva, W. Rypniewski, M. Perbandt, M. Jain, N. Genov, and C. Betzel, “Crystallization and preliminary X-ray diffraction studies of a toxic phospholipase A₂ from the venom of Vipera ammodytes meridionalis complexed to a synthetic inhibitor,” Biochimica et Biophysica Acta, vol. 1650, no. 1-2, pp. 1–3, 2003.
View at: Publisher Site | Google Scholar
D. N. Georgieva, W. Rypniewski, A. Gabdoulkhakov, N. Genov, and C. Betzel, “Asp49 phospholipase A₂-elaidoylamide complex: a new mode of inhibition,” Biochemical and Biophysical Research Communications, vol. 319, no. 4, pp. 1314–1321, 2004.
View at: Publisher Site | Google Scholar
S. L. da Silva, A. K. Calgarotto, V. Maso et al., “Molecular modeling and inhibition of phospholipase A₂ by polyhydroxy phenolic compounds,” European Journal of Medicinal Chemistry, vol. 44, no. 1, pp. 312–321, 2009.
View at: Publisher Site | Google Scholar
E. S. de Alvarenga, S. A. Silva, L. C. A. Barosa et al., “Synthesis and evaluation of sesquiterpene lactone inhibitors of phospholipase A₂ from Bothrops jararacussu,” Toxicon, vol. 57, no. 1, pp. 100–108, 2011.
View at: Publisher Site | Google Scholar

Copyright

Copyright © 2013 B. M. A. Carvalho et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

PDF Download Citation

Download other formats

Order printed copies

Views

6191

Downloads

2983

Citations