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Anti-inflammatory effects of ethanolic extract and alkamides-derived from Heliopsis longipes roots
Abstract
Heliopsis longipes (A. Gray) Blake (Asteraceae) is a broadly used species in the Mexican, Central and South American Traditional Medicine for its anaesthetic, analgesic, anti-inflammatory and anti-ulcerative properties. The ethanolic extract contains alkamides, mainly affinin (spilanthol). This family of compounds exerts an in vitro inhibitory action on the cyclooxygenase and lipoxygenase enzymes.
The present study approaches the anti-inflammatory effect of the extract and its main bioactive component affinin and derived isobutyl-decanamide.
The anti-inflammatory effect was evaluated through the mouse ear oedema test by means of two irritating agents, arachidonic acid (AA) and phorbol myristate acetate (PMA).
Heliopsis longipes, affinin and isobutyl-decanamide displayed a marked anti-inflammatory effect on the AA model with ED(50)=0.8, 1.2 and 0.9 mg/ear, respectively. Nimesulide (1 mg/ear) was used as a reference drug. In PMA model, the extract and two alkamides also showed a dose-dependent anti-inflammatory effect with ED(50)=2.0, 1.3 and 1.1 mg/ear, respectively. Indomethacin (3 mg/ear) was used as reference drug.
These results could represent an important contribution to explain the anti-inflammatory ethnobotanical effects reported for Heliopsis longipes and other species containing affinin (spilanthol). For the first time the topical anti-inflammatory effects of Heliopsis longipes, affinin and isobutyl-decanamide were studied.
... Spilanthol has many biological activities (Dubey et al., 2013), including analgesic (Molinatorres et al., 1996;Hind and Biggs, 2003;Wu et al., 2008;Cilia-López et al., 2010;Tiwari et al., 2011;Dias et al., 2012;Sharma et al., 2012;Abeysiri et al., 2013;Dubey et al., 2013;Prachayasittukal et al., 2013;Paulraj et al., 2013;Rios and Olivo, 2014;Dandin et al., 2014;Hajdu, 2014), antinociceptive (Rios et al., 2007;Déciga-Campos et al., 2012), antioxidant (Abeysiri et al., 2013), anti-inflammatory (Wu et al., 2008;Hernández et al., 2009;Dias et al., 2012), antimutagenic (Arriaga-Alba et al., 2013), antiwrinkle (Demarne and Passaro, 2009), antifungal (Dubey et al., 2013), bacteriostatic (Molina-Torres et al., 2004), insecticidal (Kadir et al., 1989;Sharma et al., 2012;Moreno et al., 2012), antimalarial (Sharma et al., 2012), anti-larvicidal activities against Aedes aegypti and Helicoverpa zea neonates (Ramsewak et al., 1999), and anti-molluscicidal activities (Johns et al., 1982). There have also been reports on its activities as an anticonvulsant, antioxidant, aphrodisiac, pancreatic lipase inhibitor, antimicrobial agent, antinociceptive agent, diuretic, vasorelaxant, anti-human immunodeficiency virus, toothache relief and as an anti-inflammatory agent (Dubey et al., 2013). ...
... Spilanthol also had an anti-inflammatory effect on the arachidonic acid model with ED 50 = 1.2 mg/ear (Wu et al., 2008). In a different study using the phorbol myristate acetate model, spilanthol showed an anti-inflammatory dose-dependent effect with ED 50 = 1.3 mg/ear (Hernández et al., 2009). ...
... It is also used as a treatment for treating toothaches, so it is called the toothache plant. Spilanthol may also have analgesic (Molinatorres et al., 1996;Hind and Biggs, 2003;Cilia-López et al., 2010;Tiwari et al., 2011;Dias et al., 2012;Sharma et al., 2012;Dubey et al., 2013;Prachayasittukal et al., 2013;Paulraj et al., 2013;Wu et al., 2008;Rios and Olivo, 2014;Dandin et al., 2014;Hajdu, 2014), antinociceptive (Rios et al., 2007;Déciga-Campos et al., 2012), antioxidant (Abeysiri et al., 2013), anti-inflammatory (Wu et al., 2008;Hernández et al., 2009;Dias et al., 2012), antimutagenic (Arriaga-Alba et al., 2013), anti-wrinkle (Demarne and Passaro, 2009), antifungal (Dubey et al., 2013), bacteriostatic (Molina-Torres et al., 2004), insecticidal (Kadir et al., 1989; ...
Spilanthol (C14H23NO, 221.339g/mol) is a bioactive compound that is found in many different plants that are used as traditional remedies throughout the world. It is present in Heliopsis longipes and several species in the genus Acmella, including A. oleracea L., also known as paracress and jambu. Its leaves and flowers have sensory properties (pungency, tingling, numbing, mouth-watering) that make it a popular spice and ingredient in several Brazilian dishes. Spilanthol can exert a variety of biological and pharmacological effects including analgesic, neuroprotective, antioxidant, antimutagenic, anti-cancer, anti-inflammatory, antimicrobial, antilarvicidal and insecticidal activities. So, the aim of this review is to present a literature review on the spilanthol that describes its occurrence, chemistry, extraction and biological activities.
... Studies have demonstrated that these compounds exhibit significant bioactivity across a broad spectrum of plant and animal pathogens, including viruses, bacteria, and fungi [5]. Additionally, they possess pharmacological properties that affect animal systems, including humans, with reported activities including anti-inflammatory, analgesic, and immunomodulatory effects [6]. Thus, it is important to further the study of the distribution and synthesis of alkamides in plants. ...
... Alkamides, particularly affinin, are known for their fungicidal, bactericidal, antiviral, and other antimicrobial activities [5,6,22]. This suggests a potential role for their accumulation in H. longipes seeds as a defense mechanism against microbial pathogens. ...
The alkamide content and specific tissue localization in the cypselae of Heliopsis longipes were investigated using gas chromatography–electron ionization mass spectrometry (GC-EIMS) and multiphoton fluorescence microscopy (MPFM). GC-EIMS analysis identified two olefinic alkamides: affinin (spilanthol) and N-2-methylbutyl-2E,6Z,8E-decatrienamide. Microscopic analysis revealed that alkamides are localized within the cotyledons, and specifically compartmentalized in lipid bodies, highlighting their spatial organization. The linear unmixing of fluorescence emission fingerprints showed that affinin exhibits autofluorescence at 693 nm, corresponding to the red spectral region. This emission is attributed to the conjugated double bonds in its acyl chain. This study is the first to report on the presence and precise localization of alkamides in the cypselae of H. longipes.
... The main phytochemical Affinin and different organic extracts of H. longipes roots are responsible for observed biological effects such as local anesthetic, flavoring, insecticidal and larvicidal [16], antimicrobial, bactericidal, and fungistatic effects [17,18]. The pharmacological effects of affinin were evidenced as an anti-inflammatory in a macrophage cell line [19] and in rodents [20], antinociceptive in mice [21,22], anxiolytic and diuretic in mice [23], and antihyperalgesic [24]. Safety tests evaluated the acute toxicity in mice, and the mean lethal dose (LD50 = 113 mg/kg) was significantly higher compared to the dose required to obtain antinociception. ...
... For the first time, the present study shows that affinin and HLEE exhibited proangiogenic effects. Heliopsis longipes roots were widely studied for medicinal purposes, where a variety of extraction processes and analytical methods were used to study the principal phytochemicals, alkamides [20][21][22]24,25,42,43]. Regarding the extraction process, maceration in combination with UAE enhanced the HLEE yield threefold (54.7 g/kg roots dry weight), with respect to previous reports that used maceration extraction alone (17 g/kg roots dry weight) [29]. ...
Angiogenesis, the formation of new blood vessels, underlies tissue development and repair. Some medicinal plant-derived compounds can modulate the angiogenic response. Heliopsis longipes, a Mexican medicinal plant, is widely used because of its effects on pain and inflammation. The main bioactive phytochemicals from H. longipes roots are alkamides, where affinin is the most abundant. Scientific studies show various medical effects of organic extracts of H. longipes roots and affinin that share some molecular pathways with the angiogenesis process, with the vasodilation mechanism of action being the most recent. This study investigates whether pure affinin and the ethanolic extract from Heliopsis longipes roots (HLEE) promote angiogenesis. Using the aortic ring rat assay (ex vivo method) and the direct in vivo angiogenesis assay, where angioreactors were implanted in CD1 female mice, showed that affinin and the HLEE increased vascular growth in a dose-dependent manner in both bioassays. This is the first study showing the proangiogenic effect of H. longipes. Further studies should focus on the mechanism of action and its possible therapeutic use in diseases characterized by insufficient angiogenesis.
... Compounds involved in vasorelaxant activity seem to be pentacyclic triterpenoids, oleanolic acid and erythrodiol [35]. Spilanthol has many biological activities [37], including analgesic [7,12,19,25,26,31,32,[38][39][40][41][42][43], antinociceptive [28,44], antioxidant [19], anti-inflammatory [25,32,45], antiwrinkle [15,46]. Dias et al. (2012) studied the influence of different extraction methods and solvents, as well as different parts of the plant (flowers, leaves and stems) on the composition, and consequently on the bioactivity of the single extract [25]. ...
... In this experimental animal model it is hypothesized that spilanthol could inactivate NF-kB, down regulating the production of proinflammatory mediators. An anti-inflammatory effect of Spilanthol has also been demonstrated on the arachidonic acid model with ED50 = 1.2 mg/ear [32] as well as a dose-dependent antiinflammatory effect with phorbol myristate acetate model with ED50 = 1.3 mg/ear [45]. ...
Despite advances in medicine and numerous agents that counteract pain, millions of patients continue to suffer. Attention has been given to identify novel botanical interventions that produce analgesia by interacting with nociceptive-transducing channels. The aim of this review is to provide an overview of the actual knowledge of Acmella oleracea (L.) and its activities, particularly those that are anti-inflammatory, anti-oxidant, and painkiller. These activities are attributed to numerous bioactive compounds, such as phytosterols, phenolic compounds and N-alkylamides (spilanthol, responsible for many activities, primarily anesthetic). This review includes 99 eligible studies to consider the anti-inflammatory, anti-oxidant, and painkiller of Acmella. Studies reported in this review confirmed anti-inflammatory and anti-oxidant activities of Acmella, postulating that transcription factors of the nuclear factor-κB family (NF-κB) trigger the transcription iNOS and COX-2 and several other pro-inflammatory mediators, such as IL-6, IL-1β, and TNF-α. The antinociceptive effects has been demonstrated and have been related to different processes, including inhibition of prostaglandin synthesis, activation of opioidergic, serotoninergic and GABAergic systems, and anesthetic activity through blockage of voltage-gated Na Channels. acmella oleracea represents a promise for pain management, particularly in chronic degenerative diseases, where pain is a significant critical issue.
... In addition, several pharmacological studies have demonstrated that affinin displays analgesic (ED 50 = 1 mg/kg intraperitoneal (i.p.) in mice) [5,16], antinociceptive (ED 50 = 6.98 mg/kg per os (p.o.); ED 50 = 36 ± 5 mg/kg i.p. in mice) [6,26], anti-inflammatory (90-180 µM in macrophage cell line) [18], anxiolytic (3-30 mg/kg i.p. in mice) [6], and diuretic (800 mg/kg p.o. in mice) [27] properties. Some of these pharmacological activities have been also reported for crude organic extracts of H. longipes roots [5,6,26,[28][29][30][31]. ...
... H. longipes roots have a long tradition of culinary and medicinal use in Mexico. A number of studies have evidenced that organic extracts obtained from H. longipes roots and affinin, their major component, possess interesting biological and pharmacological activities [4][5][6]8,9,16,25,26,[28][29][30][31]58]. However, currently, no investigation has been directed toward examining the effect of H. longipes root extracts and affinin on the vascular tone. ...
Heliopsis longipes roots have been widely used in Mexican traditional medicine to relieve pain, mainly, toothaches. Previous studies have shown that affinin, the major alkamide of these roots, induces potent antinociceptive and anti-inflammatory activities. However, the effect of H. longipes root extracts and affinin on the cardiovascular system have not been investigated so far. In the present study, we demonstrated that the dichloromethane and ethanolic extracts of H. longipes roots, and affinin, isolated from these roots, produce a concentration-dependent vasodilation of rat aorta. Affinin-induced vasorelaxation was partly dependent on the presence of endothelium and was significantly blocked in the presence of inhibitors of NO, H2S, and CO synthesis (NG-nitro-l-arginine methyl ester (l-NAME), dl-propargylglycine (PAG), and chromium mesoporphyrin (CrMP), respectively); K+ channel blockers (glibenclamide (Gli) and tetraethyl ammonium (TEA)), and guanylate cyclase and cyclooxygenase inhibitors (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) and indomethacin (INDO), respectively). Our results demonstrate, for the first time, that affinin induces vasodilation by mechanisms that involve gasotransmitters, and prostacyclin signaling pathways. These findings indicate that this natural alkamide has therapeutic potential in the treatment of cardiovascular diseases.
... In traditional medicine, chilcuague root is used to treat respiratory diseases, as an anesthetic to treat tooth and muscular aches, to treat buccal lesions, and as an anti-parasitic (Cilia López, 2007). Several alkamides have been isolated from H. longipes, but affinin, also known as spilanthol ((2E,6Z,8E)-N-isobutyl-2,6,8decatrienamide), was identified as the main alkamide in the plant (Molina-Torres et al., 1996;García-Chávez et al., 2004;Hernández et al., 2009). The oily substance causes intense lip numbness and tingling within a few minutes after placed in the mouth, and it also stimulates salivation (Correa et al., 1971). ...
... With regard to anti-inflammatory activity, Hernández et al. (2009) evaluated H. longipes ethanolic extract and purified affinin on mouse ear edema induced by either arachidonic acid (AA) or phorbol 12-myristate 13-acetate (PMA), demonstrating a significant anti-inflammatory effect. DE 50 values of 0.8 mg/ear and 1.2 mg/ear were obtained, respectively, in the AA-induced edema model; nimesulide (1 mg/ear) was used as reference drug. ...
This study assesses the anti-arthritic effect of the affinin-enriched (spilanthol, main alkamide) hexane extract from the roots of Heliopsis longipes (A. Gray) S.F. Blake, Asteraceae, on a Freund adjuvant-induced arthritis model in rodents. The extract was orally administered at a dose of 2, 6.6, or 20 mg/kg; a significant edema-inhibitory activity in the acute and chronic phases was observed with a dose of 2 and 20 mg/kg, respectively. The extract showed higher anti-inflammatory and anti-arthritic effects than the reference drug phenylbutazone (80 mg/kg). Moreover, the extract prevented the occurrence of secondary lesions associated to this pharmacological model.
... Among the vast diversity of natural products attractive for this purpose, affinin (also known as spilanthol) is a bioactive natural compound that stands out among all because it possesses the biological activity of a powerful oral anesthetic (Molina-Torres et al. 1999;Déciga-Campos et al. 2010), analgesic (Rios et al. 2007;Cariño-Cortés et al. 2010), anti-inflammatory, antimicrobial and antimutagenic agent (Molina-Torres et al. 1999;Hernández et al. 2009;Arriaga-Alba et al. 2013). Affinin IUPAC name is (2E, 6Z, 8E)-N-isobutyl-2,6,8-decatrienamide (Molina-Torres et al. 1996). ...
... On the other hand, it has been shown that CTS nanofibers are effective in inhibiting the growth of different bacteria (Jung et al. 2007;Spasova et al. 2008;Son et al. 2009), and the antibacterial activities of affinin and AgNP have also been previously reported (Hernández et al. 2009;Kemp et al. 2009). Therefore, it is expected that the combination of these three components could produce a synergistic effect to give rise to a composite material with potentiated antibacterial activity. ...
A novel nanofibrous chitosan-based composite containing affinin and silver nanoparticles is obtained by electrospinning. Silver nanoparticles are synthesized by sunlight photoreduction of the metal complex [Ag2–(affinin)](NO3)2 in polymeric solution, via a green one-pot methodology, wherein chitosan and affinin act as reducing, dispersing and stabilizing agent.
... Especially for Echinaceae NAAs, the anti-inflammatory and immuno-modulatory properties have been well investigated, confirmed and patented (Woelkart and Bauer, 2007; Esanu, 1981). However, also the, immune effects of Anacyclus pyrethrum and antiinflammatory effects of Spilanthes, Heliopsis, Piper and Achillea species are established (Hernandez et al., 2009; Mullerjakic et al., 1994; Rimbau et al., 1996; StöhrSt¨Stöhr et al., 1999; Wu et al., 2008). Different targets for the anti-inflammatory properties have been identified. ...
... Different targets for the anti-inflammatory properties have been identified. Spilanthol inactivates NF-kB, shows significant topical anti-inflammatory effects in the mouse ear edema test and is the only NAA that has a demonstrated influence on the transcription and translation of the COX enzymes (Hernandez et al., 2009; Wu et al., 2008). All other investigated NAAs have no influence on transcriptional nor translational level of COX (Hinz et al., 2007), but their enzyme activity decreases in the presence of low concentrations of NAAs containing multiple alkyne groups in their fatty acid chain (Clifford et al., 2002 ). ...
N-Alkylamides (NAAs) are a promising group of bioactive compounds, which are anticipated to act as important lead compounds for plant protection and biocidal products, functional food, cosmeceuticals and drugs in the next decennia. These molecules, currently found in more than 25 plant families and with a wide structural diversity, exert a variety of biological-pharmacological effects and are of high ethnopharmacological importance. However, information is scattered in literature, with different, often unstandardized, pharmacological methodologies being used. Therefore, a comprehensive NAA database (acronym: Alkamid) was constructed to collect the available structural and functional NAA data, linked to their occurrence in plants (family, tribe, species, genus).
For loading information in the database, literature data was gathered over the period 1950-2010, by using several search engines. In order to represent the collected information about NAAs, the plants in which they occur and the functionalities for which they have been examined, a relational database is constructed and implemented on a MySQL back-end.
The database is supported by describing the NAA plant-, functional- and chemical-space. The chemical space includes a NAA classification, according to their fatty acid and amine structures.
The Alkamid database (publicly available on the website http://alkamid.ugent.be/) is not only a central information point, but can also function as a useful tool to prioritize the NAA choice in the evaluation of their functionality, to perform data mining leading to quantitative structure-property relationships (QSPRs), functionality comparisons, clustering, plant biochemistry and taxonomic evaluations.
... The control group received saline as treatment, once a day, and consecutive 5 Inflammation occurred after 30 min, and the animals were sacrificed through cervical dislocation. A 6 mm section from each ear was removed using a metal punch and then weighed [18]. Ear edema was determined by the difference in weights of the right and the left ears and expressed as edema weight, using Eq. ...
... The statistical significance of differences between the control and test groups was determined by means of one-way analysis of variance (ANOVA), followed by unpaired student's t-test. P values less than 0.05 were considered significant [18]. ...
The purpose of this study was to improve the dissolution rate and anti-inflammatory effect of ibuprofen by a solid dispersion (SD) method. Initial screening was developed based on drug solubility in carriers in the liquid state to select a suitable water-soluble carrier system for the preparation of SDs. The dissolution of ibuprofen in urea was higher than in PEG4000 or mannitol. Thus, urea was selected as the carrier for the preparation of SDs. SDs were characterized in terms of dissolution, differential scanning calorimetry (DSC), X-ray diffraction (XRD), scanning electron microscopy (SEM), and Fourier transform infrared (FTIR) spectroscopy. Solid dispersion-based (SDBT) and conventional (CT) tablets were prepared by the wet granulation method. The anti-inflammatory effect of SDBT was evaluated using the mouse ear edema test with xylene. In vitro release results indicated that the ibuprofen dissolution rate was improved by the SD. SD characterization results suggested that ibuprofen partly precipitates in crystalline and amorphous forms after SD preparation and that ibuprofen and urea do not interact. SDBT displayed more significant anti-inflammatory effects than CT. The dissolution rate and anti-inflammatory effect of ibuprofen were significantly enhanced by the ibuprofen-urea SD.
... The roots of H. longipes have traditional uses, such as condiment, medicinal and insecticide (Cilia-López et al. 2008;Parola-Contreras et al. 2020b). For several decades, the roots have been objects of many chemical researches (García-Chávez et al. 2004;Parola-Contreras et al. 2020a), and pharmacologic studies (Molina-Torres et al. 1999;Ramírez-Chávez et al. 2000;Rios et al. 2007;Hernández et al. 2009;Cariño-Cortés et al. 2010;Cilia-López et al. 2010Arriaga-Alba et al. 2013;Castro-Ruiz et al. 2017;Escobedo-Martínez et al. 2017). All these efforts have focused on to increase the knowledge to be used for industrial exploitation. ...
Heliopsis longipes is an endemic species from Sierra Gorda, México. Its collects imply the whole plant destruction due to people only use the root, which contains affinin, a pharmacologically important compound. Thus, H. longipes has been considered as a species in danger of extinction, due to unsustainable exploitation to obtain their roots for affinin commercialization. Moreover, no studies on the genetic diversity of H. longipes have been carried out. Taking together, these aspects suggest that it is urgent to develop strategies for sustainable production and exploitation of H. longipes. Based on the aforementioned, this study aimed to evaluate the effect of the greenhouse acclimation time and the controlled elicitation on morphological, and biochemical variables as well as the affinin content in populations of H. longipes from Central Mexico characterized in genetic diversity. Biochemical variables associated with plant stress response like superoxide dismutase (SOD), catalase (CAT) and phenylalanine ammonia-lyase (PAL) activities were measured. Moreover, affinin content was quantified by HPLC and the activity of valine decarboxylase (VDC)-associated to affinin contents was also determined. Additionally, genetic variability of H. longipes populations evaluated using DNA markers was also analyzed to determine genetic diversity of accessions before greenhouse studies. Results on genetic diversity were calculated in the populations from Conca (0.27), SJB (0.15) and Beltran (0.11). The average genetic diversity was 0.21, which places this species in the genetic diversity in danger of extinction. Because of the low genetic diversity found among populations of H. longipes in Central México regions, suggests that any one of the H. longipes populations studied might be used for acclimation in a greenhouse to be used in controlled elicitation studies. Controlled elicitation with 200 mM H2O2 dose significantly increased PAL and VDC enzymatic activities as well as the affinin content in the roots of H. longipes at 7-months of acclimation time in greenhouse. Our results displayed that it was possible to increase affinin content after 7-months acclimation in greenhouse in H. longipes accessions from Central México. Thus also contributing to sustainable exploitation of this species and to decrease its danger of extinction for overexploitation.
... g/ mol) (Molina-Torres et al. 1996; Barbosa et al. 2015). Spilantol has been used in scientific studies to demonstrate its relationship with several biological effects, such as analgesic, neuroprotective, anticonvulsant, antioxidant, and anti-inflammatory effects (Wu et al. 2008;Hernández et al. 2009;Dias et al. 2011;Silva and Oliveria 2013). ...
Fatty amides (N-alkylamides) are bioactive lipids that are widely distributed in microorganisms, animals, and plants. The low yield in the extraction process of spilantol, a fatty amide, which is mainly related to its diverse biological effects, compromises its application on a large scale. Thus, this study proposes an alternative method to synthesise fatty amides from Bertholletia excelsa (AGBe) oil, with a chemical structure similar to that of spilantol. Carrageenan-induced abdominal oedema in vivo models were used in zebrafish (Danio rerio). In in vivo studies, oral AGBe produced no signs of toxicity. In the histopathological study, AGBe did not cause significant changes in the main metabolising organs (liver, kidneys, and intestines). All doses of AGBe (100 mg/kg, 500 mg/kg, and 750 mg/kg) were effective in reducing oedema by 65%, 69%, and 95%, respectively, producing a dose–response effect compared to the control group, and spilantol-inhibited oedema by 48%. In the in silico study, with the use of molecular docking, it was observed that among the AGBe, the molecules 18:1, ω-7-ethanolamine, and 18:1, ω-9-ethanolamine stood out, with 21 interactions for COX-2 and 20 interactions for PLA2, respectively, surpassing the spilantol standard with 15 interactions for COX-2 and PLA2. The anti-inflammatory action hypothesis was confirmed in the in silico study, demonstrating the involvement of AGBe in the process of inhibiting the enzymes COX-2 and PLA2. Therefore, based on all the results obtained and the fact that until the dose of 1000 mg/kg was administered orally in zebrafish, it was not possible to determine the LD50; it can be said that AGBe is effective and safe for anti-inflammatory activity.
Graphic abstract
... Heliopsis longipes (commonly known as chilcuague) is a perennial herbaceous plant endemic of Sierra Gorda, comprising regions of the states of Guanajuato, Queretaro and San Luis Potosí in the center of Mexico (García-Chávez et al. 2004). This species belongs to the Asteraceae family and is the species with the most economic importance of its genus, as its root is widely used by the traditional Mexican, Central and South American medicine for its anesthetic, analgesic, antiulcer, anti-inflammatory and antiparasitic properties; furthermore it is also used as a culinary seasoning and insecticide (Hernández et al. 2009). Up to date, the studies realized with extracts obtained from the H. longipes root, have demonstrated that the diverse pharmacological effects are due to the presence of a bioactive alkamide, called affinin (Escobedo-Martínez et al. 2017, Willig et al. 2019. ...
Background:
Heliopsis longipes root is the widely used organ from the plant due to its culinary and medicinal importance. Nevertheless, during its collection the foliage is discarded. Therefore, it is required to perform studies about the phytochemical content with biological activity present in the different plant organs of the foliage, for a better exploitation of the plant.
Hypothesis:
The concentration of phenolic compounds with antioxidant activity varies according to the type of plant organ and the geographic location of the plant collection.
Study species:
Heliopsis longipes
Site and years of study:
The H. longipes plants were collected in three locations (Conca, San Juan Buenaventura and Beltran) from Sierra Gorda of Mexico, between 2017 and 2018.
Methods:
In this study, they were analyzed the methanolic extracts from H. longipes leaves and flowers, in order to determine the content of the phenolic compounds and scavenging of radicals activity (DPPH• and ABTS•+). Further characterization of the extracts was carried out by chromatography (UPLC).
Results:
The total content of polyphenols, flavonoids and condensed tannins varied in depending on the type of plant organ and the origin site of H. longipes, being the leaves of Beltran those that represent the best source of phenolic compounds with scavenging activity of the radicals DPPH• and ABTS•+. In the extracts, phenolic acids and flavonoids were identified.
Conclusions:
The geographical climatic conditions influence the phenolic content and antioxidant activity of the methanolic extracts from leaves and flowers of H. longipes.
... The in vivo anti-inflammatory activity of the flower extract can be attributed to the main presence of spilanthol, responsible for most of the pharmacological activities described for the related species (Freitas-Blanco et al., 2016). Similarly, spilanthol and an ethanolic extract of Heliopsis longipes (Asteraceae) rich in alkamides were evaluated in a mouse ear edema model, demonstrating the topical anti-inflammatory activity of spilanthol with dose-dependent effects that were superior compared to the extract (Hernández et al., 2009). A study applying an in vivo formalin-induced hyperalgesia model revealed that a cold-water extract of S. acmella attenuated inflammatory pain at the initial and late phases in a range of doses of 500-1500 mg/kg, possibly inhibiting prostaglandin synthesis (Ratnasooriya and Pieris, 2005). ...
Ethnopharmacological relevance
Acmella oleracea (L.) R. K. Jansen (Asteraceae), known as jambú in Brazil, is used in traditional medicine as analgesic and for inflammatory conditions, characterized by the presence of N-alkylamides, mainly spilanthol. This bioactive compound is responsible for the above-described pharmacological properties, including sialagogue and anesthetic.
Aim of the study
This study aimed to characterize the anti-inflammatory effects of A. oleracea leaves (AOEE-L) and flowers (AOEE-F) extracts, including an isolated alkylamide (spilanthol), using in vitro and in vivo models. The mechanism underlying this effect was also investigated.
Materials and methods
Extracts were analyzed by HPLC-ESI-MS/MS in order to characterize the N-alkylamides content. AOEE-L, AOEE-F (25-100 μg/mL) and spilanthol (50-200 μM) were tested in vitro on VSMC after stimulation with hyperglycemic medium (25 mM glucose). Their effects over nitric oxide (NO) generation, chymase inhibition and expression, catalase (CAT), superoxide anion (SOD) radical activity were evaluated. After an acute administration of extracts (10-100 μg/mL) and spilanthol (6.2 μg/mL), the anti-inflammatory effects were evaluated by applying the formalin test in rats. Blood was collected to measure serum aminotransferases activities, NO activity, creatinine and urea.
Results
A number of distinct N-alkylamides were detected and quantified in AOEE-L and AOEE-F. Spilanthol was identified in both extracts and selected for experimental tests. Hyperglycemic stimulation in VSMC promoted the expression of inflammatory parameters, including chymase, NO, CAT and SOD activity and chymase expression, all of them attenuated by the presence of the extracts and spilanthol. The administration of extracts or spilanthol significantly inhibited edema formation, NO production and cell tissue infiltration in the formalin test, without causing kidney and liver toxicity.
Conclusion
Taken together, these results provide evidence for the anti-inflammatory activity of leaves and flowers extracts of jambú associated distinctly with their chemical profile. The effects appear to be associated with the inhibition of chymase activity, suppression of the proinflammatory cytokine NO and antioxidant activities.
... Alkylamides have also been reported to inhibit both COX-1 and of COX-2 to a different extent with COX-1 inhibition highest for undeca-2Z,4E-diene-8,10-diynoic acid isobutylamide, dodeca-2E,4Z-diene-8,10-diynoic acid 2-methylbutylamide, and undeca-2E,4Z-diene-8,10-diynoic acid 2-methylbutylamide at concentrations of 100 μg/ml, while undeca-2Z,4E-diene-8,10-diynoic acid isobutylamide had the highest COX-2 inhibition (Müller-Jakic et al. 1994;Clifford et al. 2002). The antiinflammatory assessment of alkylamides from H. longipes evaluated in arachidonic acid (AA)-and phorbol myristate acetate (PMA)-induced mouse ear edema showed that affinin and isobutyl-decanamide had a good anti-inflammatory effect with ED 50 = 0.8, 1.2, and 0.9 mg/ear, respectively, in the AA model as well as a dose-dependent anti-inflammatory effect with the extract, affinin, and isobutyl-decanamide having ED 50 of 2.0, 1.3, and 1.1 mg/ear, respectively, in the PMA model (Hernández et al. 2009). Other anti-inflammatoryrelated reports include antiseptic effects of pellitorine in HMGB1-induced inflammatory responses and its vascular barrier protective effects in LPS-induced inflammation . ...
Interest in alkylamide, as a class of compound, has grown tremendously in recent years. This interest is due to the many presumed benefits in food, cosmetics, and medicine. This review focuses on the different alkylamides naturally occurring in many plant species. Several methods have been employed for their identification as well as isolation and several in vitro and in vivo studies have revealed their therapeutic effects in various diseases. In general, alkylamides have been reported to have several biological activities and pharmacological effects which include immunomodulatory, antithrombotic, antimicrobial, antiviral, antioxidant, anti-inflammatory, analgesic, anticancer, antidiabetic, and antiprotozoal activities. Moreover, many studies have reported on their mechanisms of action, structure-activity relationship, pharmacokinetics, and toxicity. We herein present an updated report on the chemistry and pharmacology of alkylamides of natural origin.
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... Heliopsis longipes (commonly known as chilcuague) is a perennial herbaceous plant endemic of Sierra Gorda, comprising regions of the states of Guanajuato, Queretaro and San Luis Potosí in the center of Mexico (García-Chávez et al. 2004). This species belongs to the Asteraceae family and is the species with the most economic importance of its genus, as its root is widely used by the traditional Mexican, Central and South American medicine for its anesthetic, analgesic, antiulcer, anti-inflammatory and antiparasitic properties; furthermore it is also used as a culinary seasoning and insecticide (Hernández et al. 2009). Up to date, the studies realized with extracts obtained from the H. longipes root, have demonstrated that the diverse pharmacological effects are due to the presence of a bioactive alkamide, called affinin (Escobedo-Martínez et al. 2017, Willig et al. 2019. ...
Phenolic compounds and antioxidant activity of methanolic extracts from leaves and flowers of chilcuague (Heliopsis longipes, Asteraceae) Compuestos fenólicos y actividad antioxidante de extractos metanólicos de hojas y flores de chilcuague (Heliopsis longipes, Asteraceae)
... The results of this analysis showed that the pharmacological activity of A. oleracea species is due to spilanthol, which, according to Dubey et al. [31], has many pharmacological properties, of which the anesthetic activity is the most prominent [32][33][34][35]: it also have antioxidant [36], anti-inflammatory [37], anti-wrinkle [38], antifungals [31], antimalarials [35] activities. There are also reports indicating anticonvulsant, antioxidant, aphrodisiac, pancreatic lipase inhibitor, antimicrobial, antinociceptive, diuretic, vasorelaxant [31] properties. ...
Hydroethanolic preparations of the botanical species Acmella oleracea L. are used in the north of Brazil for the treatment of various diseases. However, few studies have been conducted to evaluate the toxicity of this species. The objective of this study was to evaluate the acute toxicity of the hydroethanolic extract of A. oleracea L. (EHFAo) flowers in zebrafish by immersion and oral administration. The extract was analyzed by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). EHFAo was administered orally (44.457, 88.915, 199.94, 281.83, and 448.81 mg/kg) and by immersion (250, 300, 350, 400, and 450 µg/L). Behavioral and histopathological analysis of gills, liver, intestine, and kidney were performed. The presence of (2E,6Z,8E)-N-isobutyl-2,6,8-decatrienamide (spilanthol) in EHFAo was identified by ultra-high-re.solution liquid chromatography-electrospray ionization mass spectrometry (UHPLC-ESI-MS). Treatment with EHFAo caused significant behavioral changes and death. The calculated median lethal dose (LD 50) was 148.42 mg/kg, and the calculated median lethal concentration (LC 50) was 320 µg/L. In the histopathological study, it was observed that upon oral treatment, the tissue alterations that compromised the normal functioning of the organism occurred with EHFAo doses of 88.915, 199.53, and 281.83 mg/kg, the intestine being the most affected. When the treatment was performed by immersion, the most toxic EHFAo concentrations according to the histopathological evaluation were 300, 350, and 400 µg/L, with the most affected organ being the gills. Finally, EHFAo in this study was shown to be more toxic to the liver, intestine, and kidneys when administered orally and to gills, liver, and kidneys when administered by immersion in water. Therefore, considering the results obtained and the chemical characteristics of the main phytochemical marker of EHFAo, spilanthol, it can be suggested that, depending on the dose, this compound can lead to histopathological damages in the organs highlighted in this study.
... Spilanthol showed solubility in chloroform and it had its structure determined by NMR, according to the data compared in the literature. 52 Spilanthol (Figure 3) is an N-alkylamide with varied bioactivity, 52,53 including analgesic activity, 6,52,54-61 antinociceptive, 62 antioxidant, 58 anti-inflammatory, 56,63 antimutagenic, 64 anti-wrinkle, 65 antifungal, 52 bacteriostatic, 66 insecticides, 11,57 antimalarial, 57 anti-larvicidal activities against Aedes aegypti and Helicoverpa zea neonates, 18 and antimolluscicidal activities. 67 There are also reports of its activities as an anticonvulsant, antioxidant, aphrodisiac, pancreatic lipase inhibitor, antimicrobial agent, antinociceptive agent, diuretic, vasorelaxant, human immunodeficiency virus, toothache relief, and as an anti-inflammatory. ...
Hydroethanolic preparations of Acmella oleracea is used in the north of Brazil as a female aphrodisiac. Thus, the objective of this study was to evaluate the action of the hydroethanolic extract of Acmella oleracea (EHFAo) flowers (21.873 and 44.457 mg/kg) and spilanthol (3 mg/kg) administered orally on reproductive performance and effects on the embryonic development of zebrafish F1 generation. It was observed that in the groups in which males and females received EHFAo and spilanthol, the spawning was interrupted, whereas in the groups in which only the females were treated, spawning occurred during the 21 days. Thus, in the histopathological evaluation of the gonads, it was possible to observe that the percentage of mature cells in the spermatozoa and females was significantly reduced. Only the embryo groups in which parental generation was treated with EHFAo showed lethal and teratogenic effects. On the other hand, the parental groups treated with the spilanthol presented only the lethality. Spilanthol and some metabolites showed good oral availability and important toxicological properties. Thus, it is suggested that the treatment of parental generation of zebrafish with EHFAo and spilanthol caused severe changes in the gonads and on fertility. However, on the embryo, the most striking effects in the development were recorded in the groups in which the parental generation was treated with the EHFAo, while the spilanthol influenced the lethality of the embryos.
... Similarly, a cold extract of aerial parts from Acmella oleracea administrated orally also prevented oedema formation in a carrageenan model in rats (Chakraborty et al. 2004). Hernández et al. (2009) showed that an ethanolic extract rich in alkylamides from Heliopsis longipes was more potent than the purified spilanthol (mainly alkylamide in the extract) in arachidonic acid-induced ear oedema in mice. Likewise,alkylamides with 4-fluorobenzyl or benzyl radicals on the indole ring also reduced the paw oedema induced by carrageenan in rats, the alkylamide with a 4-fluorobenzyl radical being the most potent antiinflammatory molecule tested (Fouchard et al. 2001). ...
Acmella oleracea (“jambu”) is an Amazonian plant rich in alkylamides. Its flowers are widely used in folk medicine to treat toothache due to tingling, numbness, and local anaesthesia caused in the mouth. Our group previously demonstrated that the intraplantar (i.pl.) injection of an alkylamide-rich hexane fraction (HF) obtained from jambu flowers and a synthetic isobutylalkyl amide (IBA) displayed antinociceptive and anesthetic effects in acute pain models. Thus, here we evaluated the effects of HF and IBA on carrageenan-induced acute inflammation. Mice were pretreated with HF or IBA (0.01, 0.1, and 1 µg/20 µL, i.pl.) 15 min before carrageenan injection (300 µg/20 µL, i.pl.). Mechanical allodynia and paw oedema were evaluated previously (basal) and at 0.5 until 6 h following carrageenan. Both HF and IBA at 0.1 µg promoted effective and long-lasting antiallodynic and anti-oedematogenic activities until 3 and 5 h, respectively, in comparison to the different doses evaluated. At the inflammatory peak, the plantar surfaces were excised for measurement of inflammatory and oxidative stress parameters. HF and IBA (0.1 µg) reduced the myeloperoxidase activity, TNF-α and IL-1β levels, prevented the production of lipid hydroperoxides, and the decrease of antioxidant agents, namely superoxide dismutase and catalase activities, and glutathione contents. Furthermore, only HF maintained IL-10 levels and decreased PGE2 synthesis. On the basis of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, HF and IBA are devoid of antioxidant activity in vitro. Collectively, our results demonstrated the promising anti-inflammatory effect of local pretreatment with alkylamides, supporting the potential of these molecules to treat acute inflammatory pain conditions.
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... Some other fragments were detected in product ions analysis, the fragment m/z 69 can be attributed to the group isobutylnitrile (C 4 H 7 N + ), while the fragment m/z 53 can be Evidence-Based Complementary and Alternative Medicine These authors also reported a fragment m/z = 99, which could be isobutyl isocyanate (C 5 H 9 NO). Spilanthol has several pharmacological activities reported [35], among them, its local anesthetic activity is the most reported [7,[35][36][37][38][39][40][41][42]; other activities are analgesic [43], antioxidant [39], anti-inflammatory [38,44,45], antiwrinkle [46], antifungal [35], aphrodisiac [35], antimalarial [47], among others. ...
The plant species Acmella oleracea L. is used in the north of Brazil for the treatment of a range of illnesses, such as tuberculosis, flu, cough, and rheumatism and as an anti-inflammatory agent; besides, hydroethanolic formulations with this species are popularly used as a female aphrodisiac agent. However, currently, there are no studies performed evaluating its effect on embryonic development. Hence, this research aimed to evaluate the effects of the hydroethanolic extract of A. oleracea (EHFAo) on the reproductive performance (parental) and embryonic development (F1 generation) of zebrafish, at concentrations of 50, 100, and 200 í µí¼g/L. Histopathology of parental gonads after 21 days of exposure to EHFAo reveals few alterations in the ovaries and testes, not impairing the reproduction; an increase of eggs deposition was observed in animals treated with EHFAo at the highest concentrations. Nevertheless, concerning the embryonic development of F1, teratogenic effects were observed including tail deformation, cardiac and yolk edema, scoliosis, and growth retardation; these alterations were more prominent in the groups born from progenitors exposed to the highest concentrations (100 and 200 í µí¼g/L.); but only the occurrence of yolk and cardiac edema had a statistically significant difference when compared to the control group. The chromatographic analysis shows that spilanthol (affinin) was the primary compound found in the EHFAo. Hence, in silico assessment was performed to evaluate the pharmacokinetic and toxicological properties of this molecule and 37 metabolites derived from it. Overall, our data show that the treatment caused no detrimental changes in progenitors regarding their gonads or fertility but caused some potentially teratogenic activity in embryos, which may be due to the action of spilanthol's metabolites M3, M6, M7, M8, M16, M28, and M31.
... To develop therapeutic drugs for the treatment of dermatitis, an initial step may include the identification of effective anti-inflammatory agents to abrogate and suppress inflammatory mediators in keratinocytes. Spilanthol [(2E, 6Z, 8E)-N-isobutylamide-2,6,8-decatrienamide] is a bioactive compound detected in Acmella oleracea (Spilanthes acmella) and other Acmella species, including A. brachyglossa and A. ciliata (16). A. acmella exerts a variety of biological properties, including antipyretic (17,18), anti-inflammatory (19)(20)(21), analgesic (22)(23)(24) and antimicrobial activities (22)(23)(24)(25)(26). Spilanthol inhibits lipopolysaccharide-induced inflammatory responses in murine RAW 264.7 macrophages via inactivation of the NF-κB signaling pathway (19). Leaf extracts of S. acmella have exhibited immunomodulatory activity, which may be beneficial for the treatment of rheumatism (27); spilanthol enhances immune activities in influenza and respiratory infections (28). ...
Spilanthol has been reported to possess antioxidant, anti‑inflammatory, antimicrobial and antinociceptive properties. At present, the literature has reported the beneficial role of spilanthol on tumor necrosis factor‑α (TNF‑α)‑stimulated HaCaT cells. The present study investigated the effects of spilanthol on the expression of TNF‑α‑induced intercellular adhesion molecule 1 (ICAM‑1) and cyclooxygenase (COX)‑2 in the human keratinocyte cell line HaCaT. Cells were pretreated with various concentrations of spilanthol (10‑150 µM) followed by TNF‑α to induce inflammation. Pretreatment with spilanthol decreased TNF‑α‑induced COX‑2 expression by western blotting and suppressed the expression of pro‑inflammatory mediators, including interleukin (IL)‑6, IL‑8 and monocyte chemotactic protein 1 using ELISA. Spilanthol also decreased the expression of TNF‑α‑induced ICAM‑1 protein and mRNA assay by western blotting and RT‑qPCR, respectively, in addition to the monocyte adhesiveness of HaCaT cells. Furthermore, spilanthol significantly suppressed the phosphorylation of c‑Jun N‑terminal kinase (JNK), while pretreatment with spilanthol enhanced heme oxygenase (HO)‑1 protein expression by western blotting. These results demonstrated that spilanthol may exert its anti‑inflammatory activity by suppressing the TNF‑α‑induced expression of ICAM‑1, COX‑2 and pro‑inflammatory mediators by enhancing that of HO‑1, and inhibiting the activation of the phosphorylated JNK signaling pathway. It is hypothesized that spilanthol may be a natural anti‑inflammatory drug to attenuate skin inflammatory disease.
... We found that spilanthol enhanced heme oxygenase-1 (HO-1) protein expression. HO-1 can regulate the balance of antiinflammatory mediators and has antioxidant properties [29], prompting us to evaluate the anti-inflammatory molecular mechanisms underlying the effects of spilanthol. Our results showed that spilanthol significantly reduced Fig. 7. Model explaining the mechanism underlying the anti-inflammatory effects of spilanthol (SP). ...
Spilanthol a phytochemical derived from the Spilanthes acmella plant has antimicrobial, antioxidant, and anti-inflammatory properties. This study evaluated its effects on the expression of intercellular adhesion molecule 1 (ICAM-1) and inflammation-related mediators in IL-1β-stimulated human lung epithelial A549 cells. Human lung epithelial A549 cells were pretreated with various concentrations of spilanthol (3–100 μM) followed by treatment with IL-1β to induce inflammation. The protein levels of pro-inflammatory cytokines, chemokines, and prostaglandin E2 (PGE2) were measured using ELISA. Cyclooxygenase-2 (COX-2), heme oxygenase (HO-1), nuclear transcription factor kappa-B (NF-κB), and mitogen-activated protein kinase (MAPK) were measured by immunoblotting. The mRNA expression levels of ICAM-1 and MUC5AC were determined by real-time polymerase chain reaction. Spilanthol decreased the expression of PGE2, COX-2, TNF-α, and MCP-1. It also decreased ICAM-1 expression and suppressed monocyte adhesion to IL-1β-stimulated A549 cells. Spilanthol also significantly inhibited the phosphorylation of MAPK and I-κB. These results suggest that spilanthol exerts anti-inflammatory effects by inhibiting the expression of the pro-inflammatory cytokines, COX-2, and ICAM-1 by inhibiting the NF-κB and MAPK signaling pathways.
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... Multiple studies have shown that the inhibitory effects of plant extracts and NSAIDs in similar animal models of pain and inflammation [36][37][38][39]. It is known that diclofenac and aspirin inhibit inflammation and pain by attenuating prostaglandin synthesis via cyclooxygenase inhibition in arachidonic acid pathways [33]. ...
... 92a was shown to inhibit the AA-induced edema in a dose-dependent manner with an ED 50 value at 1.2 mg/ ear, while 86a displayed the same effect with an ED 50 at 0.9 mg/ear. The acute PMA-induced inflammation was inhibited with ED 50 values at 1.3 mg/ear with 92a and 1.1 mg/ear with 86a (Hernández et al. 2009). ...
... It has been widely used by Nahuatl people (the predominant indigenous group in Central Mexico) as a condiment and also in traditional medicine. For example, it is commonly employed as buccal anesthetic, analgesic for toothaches, antiulcerative, antiparasitic, anti-inflammatory (Calzada et al., 1998;Cariño-Cortes et al., 2010;Colvard et al., 2006;Hernandez et al., 2009;Ortiz et al., 2009), as well as an antimicrobial and a fungistatic agent (Molina-Torres et al., 1999). These effects have been attributed to affinin (N-isobutyl-2,6,8decatrienoamide), the most abundant and main bioactive alkamide isolated from H. longipes (García-Chávez et al., 2004;Rios, 2012). ...
... 92a was shown to inhibit the AA-induced edema in a dose-dependent manner with an ED 50 value at 1.2 mg/ ear, while 86a displayed the same effect with an ED 50 at 0.9 mg/ear. The acute PMA-induced inflammation was inhibited with ED 50 values at 1.3 mg/ear with 92a and 1.1 mg/ear with 86a (Hernández et al. 2009). ...
Alkamides are natural products formed by connecting straight-chain, mostly unsaturated, aliphatic acids with various amines by an amide linkage. More than 300 derivatives are known from eight plant families consisting of various combinations of 200 acids with 23 amines. Apart from a few saturated derivatives alkamides with unsaturated acid parts are grouped into compounds with purely olefinic patterns and those with olefinic and acetylenic linkages. Derived from C18 oleic acid the acid parts are modified either by chain elongations to C28 or by oxidative shortenings to C4 acid residues. Substrate and regiospecific desaturases and acetylenases are responsible for their characteristic patterns of unsaturation. Amine parts are derived from various amino acids by decarboxylation. Beside the widespread isobutylamines alkamides with six- and five-membered ring amines and those with phenylalanine derived amines are characteristic for the Asteraceae and Piperaceae while benzylamines are restricted to the Brassicaceae. Within the Asteraceae 2-methylbutylamine distinguishes the tribe Heliantheae from Anthemideae characterized by ring amines. Alkamides with elongated olefinic acid parts are mainly found in Piperaceae and Brassicaceae while acetylenic acid parts are typical for Asteraceae. A wide variety of biological activities ranges from the characteristic pungent/tingling property and high insecticidal toxicity to significant antifungal, antibacterial, antiprotozoal, molluscicidal, cercaricidal, and acaricidal activity. They also act as plant growth-promoting substances. Position and stereochemistry of the double bonds are essential for the different qualities of the pungent taste. Medically alkamides possess anti-inflammatory and analgesic properties and are responsible for immuno-modulatory and cannabinomimetic effects.
... It has been widely used by Nahuatl people (the predominant indigenous group in Central Mexico) as a condiment and also in traditional medicine. For example, it is commonly employed as buccal anesthetic, analgesic for toothaches, antiulcerative, antiparasitic, anti-inflammatory (Calzada et al., 1998;Cariño-Cortes et al., 2010;Colvard et al., 2006;Hernandez et al., 2009;Ortiz et al., 2009), as well as an antimicrobial and a fungistatic agent (Molina-Torres et al., 1999). These effects have been attributed to affinin (N-isobutyl-2,6,8decatrienoamide), the most abundant and main bioactive alkamide isolated from H. longipes (García-Chávez et al., 2004;Rios, 2012). ...
Heliopsis longipes (A. Gray) Blake (Asteraceae), a plant native to Mexico, is used in traditional medicine as analgesic and microbicide. The main component in the H. longipes ethanolic extract (HLEE) is affinin, as determined by HPLC/UV-visible and NMR measurement. To date, there is no documented evidence on the spermicidal activity of this extract.
The objective of this study was to assess in vitro the effectiveness of HLEE as spermicide.
The spermicidal activity of HLEE was evaluated by the Sander-Cramer assay. Spermatozoa were incubated for 20 s with HLEE in concentrations ranging from 75 to 2000 µg/mL to determine the minimum effective concentration (MEC) value. The 50% effective concentration (EC50) of HLEE was estimated by assaying serial dilutions from the MEC. Additionally, sperms were incubated with 125, 250, or 500 µg/mL of HLEE to evaluate the viability and the integrity of sperm membrane. Lipid peroxidation was assessed by the thiobarbituric acid reactive substances assay.
HLEE caused an inhibition of 100% in spermatozoa motility at a MEC value of 2000 µg/mL; the EC50 value was 125 µg/mL. Additionally, exposure to HLEE at 125, 250, or 500 µg/mL for 30 min decreased sperm viability to 27%, 8%, and 2% of the control value, respectively, and significantly increased the percentage of sperms with structurally disorganized membrane. HLEE also increased significantly the level of lipid peroxidation in sperms with respect to controls.
The results demonstrate the spermicidal activity of HLEE in vitro and suggest that this action is caused by oxidative damage and alterations in the spermatozoal membrane.
... Spilanthol also showed interesting bioactivities such as strong local anesthetic and analgesic [12,27,28] as well as insecticidal activities [29]. The topical in vitro and in vivo anti-inflammatory effects of isolated spilanthol and of reduced amide isobutyl-decanamide have been recently reported, suggesting that spilanthol attenuates the induced inflammatory responses in macrophages by regulating the production of proinflammatory mediators [12,30,31], being a potential new lead compound for non-steroidal anti-inflammatory drugs. Although anti-inflammatory drugs are used extensively, their prolonged consumption is usually associated with numerous side effects. ...
Spilanthes acmella var oleracea, commonly known as jambú, is a natural source of secondary metabolites such as alkylamides which are responsible for the anti-inflammatory, antiseptic and anesthetic bioactivities of the plant. The purpose of this work was to characterize the extracts obtained from jambú flowers, leaves and stems by a fractionated extraction procedure that included a supercritical fluid extraction (SFE) step, using supercritical carbon dioxide as solvent, followed by an enhanced solvent extraction (ESE) step using pressurized CO2 with ethanol, water and their mixtures as solvent enhancers. Results show that the flowers are richer in spilanthol, which justifies the highest antioxidant/total phenolic ratio as well as the highest anti-inflammatory activity. SFE proved to be particularly selective for spilanthol, mainly in the case of the flowers, yielding solvent free extracts with a yellow color, adequate to be used without further time consuming and solvent dependent purification processes.
... The first chemical analysis of this plant was performed by Haller in 1945 (Acree et al., 1945a,b), and in 1947, H. longipes Blake was identified as source of affinin, which previously had been thought to derive from Erigeron affinis D. C (Jacobson et al., 1947). Since its first analysis, this medicinal plant has been studied by several research groups, with the purpose of investigating the fungistatic and bacteriostatic (Molina-Torres et al., 2004), analgesic (Rios et al., 2007), anti-hyperalgesic (Acosta-Madrid et al., 2009), antinociceptive (Ogura et al., 1982; Dé ciga-Campos et al., 2010; Cariñ o-Corté s et al., 2010), and anti-inflammatory properties (Herná ndez et al., 2009). The roots from H. longipes have been used in folk medicine to treat pain, particularly tooth and oral pain (Correa et al., 1971; Jacobson, 1954), and ulcerative conditions (Correa et al., 1971; Colvard et al., 2006; Gutié rrez-Lugo et al., 1996). ...
From the fresh roots of Heliopsis longipes three new minor alkamides: longipinamide A (N-isobutyl-8,10-diynoic-3Z-undecenamide), longipenamide A (N-isobutyl-syn-8,9-dihydroxy-2E,6Z-decadienamide) and longipenamide B (N-isobutyl-syn-6,9-dihydroxy-2E,7E-decadienamide); three known alkamides: affinin (spilanthol, N-isobutyl-2E,6Z,8E-decatrienamide), N-isobutyl-2E,6Z-decadienamide and N-isobutyl-2E-decenamide; and 11 other known compounds were isolated. The structures of the three new minor alkamides were established by 1D and 2D NMR spectroscopy including 1H, 13C, DEPT, COSY, HSQC, and HMBC experiments, as well as by EI and FAB+ mass spectrometry. To our knowledge, this is the first report of the isolation of linear dihydroxyalkamides as natural products
... Affinin displays insecticidal [4], fungicidal [5], molluscicidal [6], and antimicrobial [7,8] activities, and more recently it has been reported that this alkamide is able to alter the growth and development of Arabidopsis thaliana [9]. Affinin and other alkamides from Echinacea, Achillea and Piper species exhibit in vitro inhibitory activity against 5-lipoxygenase and cyclo-oxygenase [10][11][12][13][14]. ...
Acmella radicans (Asteraceae) produces at least seven alkamides, most with either an isobutyl- or phenylethyl group as the amine moiety. These moieties suggest that the amino acids valine and phenylalanine are the biosynthetic precursors of these alkamides. On the basis of labeled feeding experiments using either L-[2H8]valine or L-[2H8]phenylalanine we present evidence for the involvement of these two amino acids in the biosynthesis of (2E,6Z,8E)-N-isobutyl-2,6,8-decatrienamide (affinin) (1), (2Z,4E)-N-(2-phenylethyl)-2,4-octadienamide (2), (2E)-N-(2-phenylethyl)-nona-2-en-6,8-diynamide (3), and 3-phenyl-N-(2-phenylethyl)-2-propenamide (4). Alkamides were isolated from young A. radicans plants and analyzed by gas chromatography-mass spectrometry (GC-MS). Additionally, in cell free in vitro experiments based on isobutyl and phenylethylamide biosynthesis, using a colorimetric assay and GC-MS, valine and phenylalanine decarboxylase activities were assayed in the soluble extract of A. radicans leaves.
... Moreover, it has been reported that a solution of dichloromethaneextract from H. longipes showed analgesic activity as determined by gamma-aminobutyric acid (GABA) release in mice brain slices 0378 ( Ríos et al., 2007), and the interaction between H. longipes ethanolic extract (HLEE) and diclofenac on the thermal hyperalgesia test suggested that low doses of HLEE-diclofenac combination could interact synergistically at systemic level (Acosta-Madrid et al., 2009). Recently, topical anti-inflammatory effects of H. longipes, affinin and isobutyl-decanamide were confirmed ( Hernández et al., 2009). Notwithstanding the above mentioned information, to date there are no published studies regarding the relation between analgesic effect and in vivo DNA damaging potential of HLEE. ...
H. longipes S.F. Blake (Asteraceae) is a Mexican plant, whose roots are traditionally used as a condiment, as a mouth anesthetic, and as an antiparasitic. Affinin is the alkamide present in higher amounts in the roots of H. longipes.
To date, there are no published studies regarding the relation between the analgesic properties, in vivo cytotoxicity, and DNA-damaging potential of H. longipes ethanol extract (HLEE).
The HLEE was chromatographically fingerprinted to validate its affinin contents. Biological evaluation was conducted in sets of 6-8 CD1(+) mice. Antinociceptive effect was evaluated using the writhing and hot-plate tests, and mutagenic and cytotoxic effects were evaluated with micronucleous test in CD1(+) mice. For histopathological studies, biological samples from liver, heart, kidneys, spleen, lung, and brain were collected and stained.
Oral administration of HLEE (3-100 mg/kg) produced a dose-dependent antinociceptive effect in both assays. In micronucleus assay, the variability in the number of micronucleated polychromatic erythrocytes (MNPE) induced, and PE/NE index, the ratio of polychromatic erythrocytes with respect to the number of normochromatic erythrocytes induced by HLEE in the evaluated schedule, were small and nonsignificant. After histopathological results, HLEE showed polioencephalomalacia with 1000 mg/kg dose.
This work provides evidence that HLEE exerts analgesic effects, with no genotoxic effects in vivo. These findings would be an important contribution to explain the use of H. longipes root as an effective analgesic in traditional medicine, and to establish for the first time the absence of genotoxic and cytotoxic effects of the root in bioactive doses in vivo.
In previous studies we demonstrated that the ethanolic extract of Heliopsis longipes roots (EEH) and its main alkamide, affinin, elicit a vasorelaxant effect through a mechanism involving activation of the gasotransmitters pathways and stimulation of CB1 receptors and TRPA1 and TRPV1 channels. However, it has not yet been demonstrated whether EEH and affinin are capable of lowering high blood pressure. Therefore, the aim of the present study was to determine the effect of oral administration of EEH and affinin on the systolic blood pressure of L-NAME-induced hypertensive rats and to explore the participation of cannabinoid receptors and TRP channels in the mechanism of action of this alkamide. Our results showed that EEH and affinin significantly lowered systolic blood pressure and induce an improvement in endothelial function, which is associated with increased serum NO levels. Inhibition of CB1 receptors by rimonabant (3 mg/kg), TRPA1 channels by HC-030031 (8 mg/kg), and TRPV1 channels by capsazepine (5 mg/kg) significantly decreased the antihypertensive effect induced by affinin, suggesting that the blood pressure lowering effect of this alkamide involves activation of CB1 receptors and TRPA1 and TRPV1 channels.
Spilanthes acmella, also identified as Akarkara (Toothache plant), is a crucial herb with immense applications both in the medical as well as dental field. Its biologically active component, Spilanthol, is believed to be responsible for an array of wields like anti-toothache, analgesic, anti-pyretic, anti-inflammatory, anti-diuretic anti-oxidant, immune-modulatory, anti-microbial, and anti-cancer agent, etc. Recent advances in the use of this plant include food, cosmetic industry, treatment of periodontal diseases and oral ulcers, toothpaste, as an intracanal medicament, and aphrodisiac. The scope of herbal remedies for treatment is expected to rise in near future. However, more experimental studies are required to ascertain this plant’s quality, efficacy, potency, etc. The scope of the current review is to discuss various uses of Spilanthes in dentistry.
Alkamides are a group of bioactive, novel, and natural secondary compounds showing broad structural variability with an important range of activities, including tingling, pungent, antioxidant, and many more activities. Plants produce multiple intracellular and systemic responses and pathways against abiotic and biotic stresses. Alkamides are helpful in increasing plant growth, root development, the architecture of the root and shoot system, and nitric oxide (NO) signal transduction pathways, and, ultimately, yield. Their chemical-signaling pathways and relations with soil, water, and minerals lead to the presentation of defense-related genes and the production of antimicrobial secondary metabolites to have a greater impact on plant physiology and metabolism. Signals produced by alkamides and NAEs are used by microorganisms for cell-to-cell communication and can be observed by plants to modulate gene expression, metabolism, and growth development. Compounds such as alkamides produced by certain growth-promoting rhizobacteria contribute to plant immunity and development.
Background
A large group of new N-alkyl-4-methyl-2,2-dioxo-1H-2λ6,1-benzothiazine-3-carboxamides was synthesized.
Objective
On the basis of molecular docking some derivatives of N-alkyl-4-methyl-2,2-dioxo-1H-2λ6,1-benzothiazine-3-carboxamides have been designed. Their preliminary structure-activity relationships (SAR) have been studied. The most rational approaches to the synthesis of the lead compounds have been developed. The most active compounds have shown high anti-inflammatory and analgesic activities.
Methods
The structure of all compounds prepared has been confirmed by the data of elemental analysis, 1H- and 13C NMR spectroscopy, and electrospray ionization liquid chromato-mass spectrometry. For rational drug-design, optimization of further pharmacological screening and prediction of possible mechanism of pharmacological action, molecular docking has been performed. For determination of activity pharmacological studies have been carried out.
Results
Pharmacological tests have determined that the transition from N-aryl(heteroaryl)alkylamides to “pure” N-alkylamides we carried out is accompanied by a significant reduction and even complete loss of anti-inflammatory effect with remaining of analgesic activity.
Conclusion
According to the studies, compounds from N-alkyl-4-methyl-2,2-dioxo-1H-2λ6,1-benzothiazine-3-carboxamides are potential anti-inflammatory and analgesic agents.
Toothache is one of the most common global health problems, and medicinal plants are widely used to relieve the associated pain and inflammation. Several studies have been conducted on the use of plants to treat toothache, but no study has comprehensively assessed the types of plants and the mechanisms of action of the phytochemical compounds involved in their analgesic effect. This review aims to bridge this gap. This is the first review to collect a large volume of data on the global use of medicinal plants used in the treatment of toothache. It presents the relevant information for dentists, researchers, and academics on using medicinal plants to treat toothache. We found that preclinical studies and state-of-the-art technology hold promise for furthering our knowledge of this important topic. In total, 21 species of medicinal plants used to treat toothache were found in America, 29 in Europe, 192 in Africa, 112 in Asia, and 10 in Oceania. The most common species were Allium sativum, Allium cepa, Acmella oleracea, Jatropha curcas, Jatropha gossypiifolia, and Syzygium aromaticum. The most commonly found family of medicinal plants was Asteraceae, followed by Solanaceae, Fabaceae, Lamiaceae, Euphorbiaceae, Rutaceae, and Myrtaceae. The most common phytochemicals found were flavonoids, terpenes, polyphenols, and alkaloids. The reported mechanisms of action involved in toothache analgesia were antioxidant effects, effects mediated by transient receptor potential channels, the γ-aminobutyric acid mechanism, and the cyclooxygenase/lipoxygenase anti-inflammatory mechanism.
Fatty amides ( N -alkylamides) are a group of bioactive lipids widely distributed in microorganisms, animals, and plants. The low yield in the extraction process of spilantol, a grease amide, which has been related mainly to diverse biological effects, compromises its application on a large scale. Thus, this study proposed an alternative to the synthesis of fatty amides from Bertholletia excelsa (AGBe) oil, with a chemical structure similar to that of spilantol. In vivo models induced by carrageenan were used in Zebrafish ( Danio rerio ). In in vivo studies, oral AGBe produced no signs of toxicity. In the histopathological study, AGBe did not cause significant changes in the main metabolizing organs (liver, kidneys, and intestines). In the anti-inflammatory evaluation, all doses (45 mg/kg, 500 mg/kg, and 1000 mg/kg) were effective, significantly reducing edema and producing a dose-response effect when compared to spilantol. In the in silico study, with the use of molecular docking, he showed that among the AGBe, the molecules 18:1, ω-7-ethanolamine and 18:1, ω-9-ethanolamine stood out, which had 21 interactions for COX-2 and 20 interactions for PLA 2 , respectively, surpassing the spilantol standard with 15 interactions for COX-2 and PLA 2 . The hypothesis of anti-inflammatory action was confirmed in the in silico study, demonstrating the involvement of AGBe in the process of inhibiting the enzymes COX-2 and PLA 2 . Therefore, based on all the results obtained and the fact that until the dose of 1000 mg/kg, orally, in zebrafish, it was not possible to determine the LD 50 , it can be said that AGBe is effective and safe for the activity anti-inflammatory.
Background:
In Mexico, the Asteraceae are part of traditional knowledge where its members have several uses, but they are particularly remarkable in traditional medicine and are used for different purposes.
Questions:
What pharmacologically studies have been carried out with Asteraceae species used in Mexican traditional medicine? What pharmacological activities have been tested? What compounds are responsible for the tested activities?
Species studied:
Asteraceae species used in Mexican traditional medicine pharmacologically tested.
Methods:
A database including scientific studies on Asteraceae species which studies on pharmacological activity or phytochemical characterization was compiled and analyzed.
Results:
From 249 reviewed studies only 202 fulfilled the criteria for our analysis. A total of 101 species distributed in 65 genera and 16 tribes were registered. The tribes Heliantheae and Senecioneae were the most studied. Ageratina pichinchensis, Artemisia ludoviciana, Heliopsis longipes, and Heterotheca inuloides were the most studied species. In Mexico, the Asteraceae family is mainly used in the treatment of diseases or symptoms related to the digestive and respiratory systems. In 48 % of the studies some biocidal activity was evaluated but only 21.8 % included phytochemical characterizations.
Conclusions:
The antimicrobial activity and phytochemical characterizations are the main kind of ethnopharmacological studies for Asteraceae in Mexico. Most of the compounds responsible for the activities have not been identified yet. The uses of Asteraceae in Mexico are similar to other countries emphasizing its cultural importance in the world. Mexican Asteraceae should be prioritized in conservation and bioscreening schemes.
Herbal medicines are an integral element of alternative medical care in Mexico, and the best testimony to their efficacy and cultural value is their persistence in contemporary Mexican marketplaces where the highest percentages of medicinal and aromatic plants are sold. This chapter summarizes current trends in research on medicinal plants in Mexico, with emphasis on work carried out at the authors’ laboratories. The most relevant phytochemical and pharmacological profiles of a selected group of plants used widely for treating major national health problems are described.
The pandemic caused by the SARS-CoV-2 has quickly spread globally, infecting millions, and killing hundreds of thousands of people. Herein, to identify potential antiviral agents, 97 natural amide-like compounds known as alkamides and piperamides were tested against SARS-CoV-2 main protease (Mpro) and RNA-dependent RNA polymerase (RdRp), and the human angiotensin-converting enzyme 2 (ACE2) using molecular docking and molecular dynamics simulations. The docking results showed that alkamides and dimeric piperamides from Piper species have a high binding affinity and potential antiviral activity against SARS-CoV-2. The absorption, distribution, metabolism and excretion (ADME) profile and Lipinski’s rule of five showed that dimeric piperamides have druglikeness potential. The molecular dynamics results showed that pipercyclobutanamide B forms a complex with Mpro at a similar level of stability than N3-I. Our overall results indicate that alkamides and piperamides, and specifically pipercyclobutanamide B should be further studied as compounds with SARS-CoV-2 antiviral properties.
Cedrus deodara is a cedar plant from the Pinaceae family. Because Cedrus is widely used in health care products, cosmetics, medicine, and insecticides, the essential oils of Cedrus deodara (CDEOs) are gaining increasing levels of attention. However, there have only been a few studies on the anti-inflammatory effects of CDEOs. This study aimed to systematically study the composition and biological effects of CDEOs, in line with this, the study assessed the chemical composition of CDEOs and evaluated their anti-inflammatory mechanisms in mice. Gas chromatography-mass spectrometry (GC-MS) was used to determine and analyze a total of 41 compounds from CDEOs. It was found that the major components of CDEOs are thujopsene (28.23%), α-cedrene (17.01%), and cedrol (15.55%); moreover, our results showed that the active ingredients in CDEOs have anti-inflammatory and antioxidant effects and that the active ingredients in CDEOs can be used as an antioxidant, as an important biological resource in functional food, and as ingredients in cosmetics and pharmaceuticals. Taken together, these CDEOs serve as ideal additives for anti-inflammatory drugs.
Heliopsis longipes is a common spice and a medicinal plant widely used in Mexico and South America due to its mouth anesthetic, analgesic, and anti-inflammatory properties related to its spilanthol content. Previous reports have demonstrated that N-alkylamides exhibit cytotoxic effect against different cancer cell lines; however, the anti-cancer potential of the species remains unknown. Therefore, we investigated the effects of H. longipes extract (HLE) and spilanthol on HeLa, K-562, MCF-7, and HaCaT cells. HLE displayed significative antiproliferative effects and increased the rate of cell apoptosis on all cancer cell lines tested. These effects were measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell counting by flow cytometry, cell cycle analysis, Annexin V-FITC/PI activation, and nuclear DAPI staining. Significant growth inhibition was found, with emphasis to K-562 and MCF-7 cells, exhibiting IC50 values of 87.14 and 134.25 μg/mL, respectively, without inducing substantial damage to HaCaT after 48 h of treatment. Spilanthol displayed a more pronounced inhibitory effect on cell lines, maintaining the selectivity index observed for the extract. Furthermore, real-time polymerase chain reaction revealed that HLE activated caspase 3 and caspase 8 after treatment for 48 h. Annexin V-FITC staining to detect early stage of apoptosis and DAPI staining of chromatin substantiated the cytotoxic effects. GC–MS analysis of the extract confirmed spilanthol as the major constituent present (63.94%), followed by other minor alkamides, which could also contribute to the biological effect observed. These results confirm the induction of apoptosis by Heliopsis longipes as well as the potential usefulness of the species and their alkamides as compounds to develop new anti-cancer agents.
Lyophilized Acmella oleracea capitula, leaves and stems were extracted with methanol at 100oC and 10 MPa (100 atm) pressure in a sealed container and analyzed by LC-HRMS and NMR. This solubilized 28.3% of the leaves, 24.7% of the stems and 31.1% of the capitula. The NMR spectra were dominated by signals due to the CHO and CH2O of carbohydrates. Spilanthol and other N-alkamides were present in the methanolic extracts at concentrations that were too low to enable their detection by NMR. In contrast, LC-HRMS was able to quantify spilanthol (deca-2E,6Z,8E-trienoic acid isobutylamide, syn. affinin, C14H23NO) and detect at least ten other N-alkamides as [M+H]+ ions with the following molecular formulas and m/z values: C13H17NO (1), C17H17NO (2), C15H19NO (3), C15H21NO (4), C16H23NO (5), C17H23NO (6), C14H25NO (7), C15H25NO (8), Ci4H2iNO (9) and C16H25NO (10). The concentrations of spilanthol in the capitula, leaves and stems were 16.5, 0.344 and 0.241 mg/g dry weight. To confirm its presence by NMR, partial purification was required. So, a portion of the residues remaining after evaporating off the methanol were partitioned between chloroform and deionized water. The percentage of amphiphilic compounds in the capitula, leaves and stems were 7.69, 8.42 and 3.09% dry weight, respectively. NMR signals due to spilanthol were seen in their spectra.
Spilanthes acmella, a well known antitoothache plant with high medicinal usages, has been recognized as an important medicinal plant and has an increasingly high demand worldwide. From its traditional uses in health care and food, extensive phytochemical studies have been reported. This review provides an overview and general description of the plant species, bio-active metabolites and important pharmacological activities including the preparation, purification and in vitro large-scale production. Structure-activity relationships of the bioactive compounds have been discussed. Considering data from the literature, it could be demonstrated that S. acmella possesses diverse bioactive properties and immense utilization in medicine, health care, cosmetics and as health supplements. As a health food, it is enriched with high therapeutic value with high potential for further development.
The monomer N, N'-[(4, 5-dihydroxy-1, 2-phenylene)bis(methylene)]bisacrylamide (OHABA) was successfully synthesized by Friedel-Craft reaction. Using precipitation polymerization, POHABA microspheres were easily obtained in a size controlled manner. The microsphere diameters were controlled by the dosage of initiator (2，2'-azobisisobutyronitrile, AIBN). The minimum and maximum microsphere size are 0.83 ± 0.07 μm and 1.98 ± 0.13 μm, respectively. Antimicrobial activity test results showed that both OHABA and POHABA microspheres had the broad-spectrum inhibitory effect on both Gram-positive and Gram-negative bacteria, but the POHABA performed better than OHABA monomer. The inhibition rate of bigger (1.98 μm in diameter) and smaller (1.00 μm in diameter) POHABA microspheres at the concentration of 0.238 mg/mL were 74.3% and 37.0% for soil microorganisms after 48 hours reaction respectively.
Strategy, Management and Health Policy
Enabling Technology, Genomics, Proteomics
Preclinical Research
Preclinical Development Toxicology, Formulation Drug Delivery, Pharmacokinetics
Clinical Development Phases I‐III Regulatory, Quality, Manufacturing
Postmarketing Phase IV
H eliopsis longipes is a popular medicinal plant in M exico. One of the main constituents that can be extracted from H . longipes is affinin ( N ‐isobutylamide). However, available information regarding this compound is scarce, and there is only a single report related to the effect of affinin on the central nervous system. Affinin extracted from H . longipes was evaluated for its psychopharmacological activity in several models and for its safety. H . longipes extract and affinin demonstrated antinociceptive effect, modified anxiety behavior and prolonged the time of sodium pentobarbital‐induced hypnosis. Affinin elicited these activities at high doses. Both the extract and affinin decreased the time of clonic and tonic PTZ ‐induced seizures. In the A mes test, neither the extract nor affinin induced mutations in the S almonella typhimurium strains TA98 and TA100 or TA102 with or without the S9 microsomal fraction. Lethal dose 50 values in mice suggest that the H . longipes extract may contain sedative principles with potential anxiolytic activity; however, it may also increase convulsive activity.
The discovery of the interaction of plant-derived N-alkylamides (NAAs) and the mammalian endocannabinoid system (ECS) and the existence of a plant endogenous N-acylethanolamine signaling system have led to the re-evaluation of this group of compounds. Herein, the isolation of seven NAAs and the assessment of their effects on major protein targets in the ECS network are reported. Four NAAs, octadeca-2E,4E,8E,10Z,14Z-pentaene-12-ynoic acid isobutylamide (1), octadeca-2E,4E,8E,10Z,14Z-pentaene-12-ynoic acid 2'-methylbutylamide (2), hexadeca-2E,4E,9Z-triene-12,14-diynoic acid isobutylamide (3), and hexadeca-2E,4E,9,12-tetraenoic acid 2'-methylbutylamide (4), were identified from Heliopsis helianthoides var. scabra. Compounds 2-4 are new natural products, while 1 was isolated for the first time from this species. The previously described macamides, N-(3-methoxybenzyl)-(9Z,12Z,15Z)-octadecatrienamide (5), N-benzyl-(9Z,12Z,15Z)-octadecatrienamide (6), and N-benzyl-(9Z,12Z)-octadecadienamide (7), were isolated from Lepidium meyenii (Maca). N-Methylbutylamide 4 and N-benzylamide 7 showed submicromolar and selective binding affinities for the cannabinoid CB1 receptor (Ki values of 0.31 and 0.48 μM, respectively). Notably, compound 7 also exhibited weak fatty acid amide hydrolase (FAAH) inhibition (IC50 = 4 μM) and a potent inhibition of anandamide cellular uptake (IC50 = 0.67 μM) that was stronger than the inhibition obtained with the controls OMDM-2 and UCM707. The pronounced ECS polypharmacology of compound 7 highlights the potential involvement of the arachidonoyl-mimicking 9Z,12Z double-bond system in the linoleoyl group for the overall cannabimimetic action of NAAs. This study provides additional strong evidence of the endocannabinoid substrate mimicking of plant-derived NAAs and uncovers a direct and indirect cannabimimetic action of the Peruvian Maca root.
Alkamides are a group of bioactive natural compounds showing broad structural variability and an important range of biological activities, such as immunomodulatory, antimicrobial, antiviral, larvicidal, insecticidal, diuretic, pungent, analgesic, cannabimimetic, and antioxidant activities. These natural products are also involved in the potentiation of some antibiotics and the inhibition of prostaglandin biosynthesis, RNA synthesis, and the arachidonic acid metabolism. Many plant species containing alkamides have been used in traditional medicine by different civilizations around the world.
Alkamides represent a class of lipidic compounds structurally related to animal endocannabinoids. Based on the structural similarity of these compounds to anandamide (N-arachidonoylethanolamine), an endogenous cannabinoid cerebral neurotransmitter, alkamides are highly active in the central nervous system. Despite their several biological activities, their immunomodulatory and analgesic properties are most important therapeutic applications.
N-Alkylamides are a large group of bioactive molecules found in several plants from the genera Echinacea, Xanthoxylum and Spilanthes. Extracts and formulations derived from these plants are not only orally used, but also applied on the skin as well. However, there is currently no specific information available about the intrinsic local pharmacokinetics of N-alkylamides after topical application on human skin, questioning the role of this mode of administration. The present study investigates the transdermal behaviour of spilanthol, a prominent N-alkylamide.
Two pharmaceutically accepted dose solutions (ethanol and propylene glycol based aqueous donor vehicles), combined with three different receptor fluids (PBS, PBS+0.5% HPbetaCD, EtOH/H(2)O (30:70, v/v)), were applied on split-thickness human skin in a Franz diffusion cell (FDC) system. Fundamental permeation characteristics of spilanthol in a solvent-independent way (100% aqueous dose solution) were also obtained using an extrapolation approach with different organic solvent/H(2)O ratios.
We demonstrated for the first time that spilanthol permeates the skin. The following aqueous-extrapolated primary transdermal parameters were obtained (mean+/-SEM): K(p,aq)=3.31 (+/-0.29)x10(-3)cm/h, D(m,aq)=1.86 (+/-0.09)x10(-4)cm(2)/h and K(m,aq)=7.28 (+/-1.59)x10(-1). Partitioning (K(m)) was strongly dependent on the donor solution composition, while diffusion (D(m)) was mainly influenced by the receptor fluid composition.
Se describe Beaucarnea hiriartiae sp. n. del estado de Guerrero, México. Esta nueva especie es similar a B. stricta Lemaire de los estados vecinos de Puebla y Veracruz.
To examine the effects of C-phycocyanin, a pigment found in blue-green algae which acts as an antioxidant in vitro and in vivo, in different animal models of inflammation.
Male Sprague Dawley rats and OF1 mice were used.
Oedema was induced by: a) AA (0.5 mg/ear) or TPA (4 microg/ear) in the mouse ear b) carrageenan injection (0.1 mL of 1% suspension) in the rat paw (+/-adrenalectomy) and c) cotton pellet implantation in the rat axilla. Phycocyanin (50-300mg/kg, p.o.) or indomethacin (1 mg/ear or 3-10mg/kg, p.o.) as control were tested in the four animal models.
Measurement of the increase in the weight (mg) of 6 mm ear punch biopsies from treated ears were made in comparison to control ears, together with myeloperoxidase (MPO) activity as an index of neutrophil infiltration. The increase in the paw thickness (mm) was measured with a dial caliper. Cotton pellet was implanted and seven days afterwards the granuloma was removed and the dry weight was determined. Acute toxicity was studied in mice and rats. Statistics were performed using one-way analysis of variance with the Duncan Multirange test.
Phycocyanin reduced significantly (p < 0.05) and in a dose-dependent manner ear oedema induced by AA and TPA in mice as well as carrageenan-induced rat paw oedema (both in intact and adrenalectomized animals). In the TPA test, phycocyanin also reduced MPO content. Phycocyanin also exerted an inhibitory effect in the cotton pellet granuloma test. In the acute toxicity test in rats and mice, even at the highest dose tested (3000 mg/kg, p.o.), no toxicity was found.
Phycocyanin shows anti-inflammatory activity in four experimental models of inflammation. Its antioxidative and oxygen free radical scavenging properties may contribute, at least in part, to its anti-inflammatory activity.
We have investigated the immunostimulatory, anti-inflammatory, and antioxidant activities of various Echinacea raw materials and commercially available products on murine macrophages and human peripheral blood mononuclear cells (PBMCs). To emulate oral dosing, a simulated digestion protocol was employed as a means of sample preparation. Echinacea-induced macrophage activation was used as a measure of immunostimulatory activity determined via quantitative assays for macrophage-derived factors including tumor necrosis factor alpha, interleukin (IL)-1alpha, IL-1beta, IL-6, IL-10, and nitric oxide. Echinacea herb and root powders were found to stimulate murine macrophage cytokine secretion as well as to significantly enhance the viability and/or proliferation of human PBMCs in vitro. In contrast, Echinacea extracts chemically standardized to phenolic acid or echinacoside content and fresh pressed juice preparations were found to be inactive as immunostimulatory agents but did display, to varying degrees, anti-inflammatory and antioxidant properties.
Thome, Robert F. (Rancho Santa Ana Botanic Garden, Claremont, CA 91711). Classification and geography of the flowering plants. Bot. Rev.58(3): 225–348. 1992.—This treatment of the flowering plants is the latest revision of my classification of the Class Angiospermae and replaces my 1983 and more recent 1992 synopses. An update is necessary because so much new information has been published in the last decade pertinent to the classification of the flowering plants. About 870 such recent books, monographs, and other botanical papers are cited in the Introduction, listed primarily by the botanical discipline that they represent. Also considerable changes in my classification have been necessitated by my narrowed family- and ordinal-gap concepts, acceptance of the ending “-anae” for superorders in place of the traditional but inappropriate “-iflorae,” and acceptance of more prior or more widely used names for the categories above the family. A new phyletic “shrub” replaces earlier versions, and attempts to indicate visually relative sizes and relationships among the superorders, orders, and suborders. One table includes a statistical summary of floweringplant taxa: ca. 233,900 species of 12,650 genera, 437 families, and 708 subfamilies and undivided families in 28 superorders, 71 orders, and 71 suborders of Angiospermae. Three other tables summarize the known indigenous distribution of the families and subfamilies of angiosperms about the world. The synopsis lists the flowering plant taxa from the class down to the subfamily (and in Asteraceae down to the tribe) with indication of the degree of confidence I place in the circumscription and placement of each category above the subfamily, the best available estimates of the number of genera and species for each category, and the known indigenous distribution of each subfamily and family. Table V lists alphabetically the geographical abbreviations used in the synopsis. The extensive bibliography of recent literature should be helpful to those persons interested in the classification of the flowering plants.
31 crude extracts derived from 28 plants highly valued as anti-infective agents in Mexican folk medicine have been screened for antimicrobial activity against four bacteria, a yeast and two molds. The results of the quantitative study indicated that the extracts derived from five species (Malmea depressa, Heliopsis longipes, Datura lanosa, Cnidosculus tehuacanensis and Helianthella quinquenervis) possessed significant antiseptic properties, therefore supporting the ethnomedical uses of these species. The cytotoxic activity was assayed against three cell lines HT-29 (Colon adenocarcinoma), MCF-7 (Breast carcinoma), A-549 (Lung carcinoma) and only the extract of Helianthella quinquenervis possessed significant activity against the MCF-7 cell line.
imidazole} demonstrated unique antiinflammatory activities in murine models that are resistant to selective cyclooxygenase (CO) inhibitors. Both edema and inflammatory cell infiltration induced by the topical application of arachidonic acid to the mouse ear were decreased by SK&F 105809 (ed50 values of 44 mg/ kg, p.o.). Polymorphonuclear leukocyte (PMN) infiltration following the intraperitoneal injection of either monosodium urate crystal or carrageenan was inhibited with ed50 values of 64 and 72 mg/kg, p.o., respectively. These inflammatory responses were unaffected by the selective cyclooxygenase inhibitor naproxen. SK&F 105809 also inhibited leukotriene B4 (LTB4) and prostaglandin E2 production in vivo in arachidonic acid-induced inflammatory exudates (ed50 values of 41 and 15 mg/kg, p.o., respectively). The inhibition of LTB4 production preceded the inhibition of PMN infiltration. The impact of inhibition of both 5-lipoxygenase (5-LO) and CO was seen with platelet-activating factor-induced vascular permeability which was inhibited markedly by SK&F 105809. However, the 5-LO inhibitor, phenidone, only strongly inhibited when coadministered with the selective CO inhibitor, indomethacin. In spite of a short half-life (14–18 min) for both SK&F 105809 and the active metabolite imidazole}, the pharmacological activity lasted at least 1.5 hr. The biochemical evidence of inhibition of interleukin-1 (IL-1) production and 5-LO and CO activity, in vitro, by the metabolite (SK&F 105561) seen in the companion paper (Marshall PJ, Griswold DE, Breton J, Webb EF, Hillegass LM, Sarau HM, Newton J Jr, Lee JC, Bender PE and Hanna N, Pharmacology of the pyrroloimidazole, SK&F 105809—I. Inhibition of inflammatory cytokine production and of 5-lipoxygenase- and cyclooxygenase-mediated metabolism of arachidonic acid. Biochem Pharmacol42: 813–824, 1991) and inhibition of the fluid and cellular phase of the inflammatory response, in vivo, by SK&F 105809 suggest that this compound possesses a unique profile of activity.
The ability of azaspiranes to modulate the acute inflammatory response in models of skin inflammation was examined.
The in vivo experiments involved the use of 5-6 age-matched male Balb/c inbred mice (22-25 g) per treatment group and a control group of 8-10 animals. In vitro mechanistic studies used RBL-1 and U937 cells lines and freshly isolated human monocytes.
Arachidonic acid (AA) (2 mg/20 microl in acetone) or PMA (phorbol myristate acetate) (4 microg/20 microl) were applied topically. SK&F 106615 and SK&F 106610 were administered topically either dissolved in acetone or dimethylacetamide just after the application of the irritant. Isolated cells were treated with the compounds dissolved in DMSO.
The thickness and influx of neutrophils into the treated ears was measured as was the effects of the azaspiranes on 5-lipoxygenase activity, cyclooxygenase activity, prostaglandin and leukotriene synthesis, and the activation of the transcription factor NF-kappaB.
SK&F 106615 and SK&F 106610 significantly reduced inflammation in the AA- and PMA-induced inflammation models (p < 0.05) with ED50's of 179 and 120 mg/ear for edema and myeloperoxidase, respectively. The compounds did not inhibit eicosanoid biosynthesis, have a direct effect on 5-lipoxygenase or cyclooxygenase enzymes, or inhibit NF-kappaB.
The potent anti-inflammatory and immunomodulatory activities of the azaspiranes observed in these and other studies appear to be mediated by a novel mechanism.
Contributions from United States National herbarium, vol. v. published by U. S. Dept of agriculture, Division of botany vol. VIII, X, XII, XIII by Smithsonian institution, United States National museum "Ferns collected in Mexico by J. N. Rose during the months of August and September 1897. By George E. Davenport" vol. v, p. 146-151
We have found that mouse ear oedema induced by the topical application of arachidonic acid is not a specific screen for compounds inhibiting the lipoxygenase or cyclo-oxygenase pathways of arachidonic acid metabolism. Although such compounds are able to reduce the oedema substantially, pharmacological agents such as histamine antagonists, phosphodiesterase inhibitors, free radical scavengers, and also various compounds not normally considered to have anti-inflammatory properties, can equally effectively reduce the oedema. A mutual potentiation of the effects of prostaglandins, leukotrienes and mast cell-derived histamine would allow many, but not all, of the active agents to be rationalised. The ability of compounds not influencing these three types of inflammatory mediators to reduce the oedematous response means the model is of limited value for directed screening.
Structure-activity relationships obtained from in vitro screening results obviously indicate that the highest inhibition effects on cyclooxygenase and 5-lipoxygenase are found amongst the class of phenolic compounds (flavonoids, polyphenols, coumestans, phenol carboxylic acids) and arachidonic acid analogous (alkylamides, retinoids, arylheptanoids, thiosulfinates, sulfinyl disulfides). The antiinflammatory activities of some triterpenenic acids, sesquiterpene lactones, and polysaccharides may be due to their immunomodulating activities on the complement and/or T-lymphocyte populations, respectively. In the search for potential antiallergic and antiasthmatic compounds, the thiosulfinates of onion were found to be active principles of the drug. The mechanism of action of some other antiallergic plant drugs (i.e. Tylophora asthmatica, Adhatoda vasica, etc.) has not yet been clarified.
We and others have shown that arachidonic acid (AA), when applied topically to ear surfaces, causes an intense acute inflammatory reaction within minutes (as measured by ear thickness). In this study, we have investigated the cellular and biochemical changes associated with this phenomenon and have attempted to correlate these changes with the induction of inflammation. Measurement of vascular permeability by the accumulation of [125I]albumin showed that significant plasma exudation was observed at 15 min in AA-treated ears. Furthermore, the increase in [125I]albumin was time related and was nearly 10-fold greater than control at 1 h. No time-related change in plasma exudation was observed with control ears. Measurement of LTC4 by radioimmunoassay showed that there was a significant increase in LTC4 synthesis at 15 min after AA treatment. Maximal LTC4 synthesis occurred at 15 min and subsequently decreased to 30% of peak level at 30 min. Histological examination and myeloperoxidase measurement indicated that few neutrophils were present at these early time points and suggested that cells other than neutrophils are contributing to LTC4 synthesis. Ear thickness, [125I]albumin accumulation and leukotriene C4 (LTC4) synthesis in AA-treated ears were reduced significantly by topically administered mixed lipoxygenase (LO) and cyclooxygenase inhibitors such as BW755C and phenidone. Therefore, we suggest that AA-induced ear inflammation is a suitable screen for detecting LO inhibitors in vivo.
The inflammatory response of the mouse ear to topical application of arachidonic acid (2 mg/ear) was examined to study the roles of sulfidopeptide-leukotrienes (LTs) and prostaglandin (PG) E2 as mediators of edema. The increase in ear thickness caused by arachidonic acid (AA) (edema), reached a maximum at 45 to 60 min after AA application. The amounts of immunoreactive LTC4 and immunoreactive PGE2 produced increased significantly in 5 to 10 min, and then diminished gradually over 60 min. 5-lipoxygenase inhibitors, dual cyclooxygenase/lipoxygenase inhibitors and anti-histamines significantly inhibited AA-induced ear edema. Both production of PGE2 and LTC4 were suppressed by NDGA at 1 mg/ear which also inhibited ear swelling. However aspirin, which enhanced LTC4 production in AA-induced ear edema did not inhibit the ear swelling. Hypodermic injection of LTC4 at 25 ng or PGE2 at 500 ng/ear did not cause swelling, but edema was induced when both compounds were injected simultaneously. Moreover ear swelling was induced by injection of both LTD4 at 50 ng and PGE2 at 500 ng/ear. Furthermore, concomitant injection of histamine, at 500 ng or serotonin at 50 ng/ear with LTC4 at 25 ng caused ear swelling but both compounds at the same dose alone did not induce swelling. These results suggest that AA-induced ear edema is predominantly mediated by LTC4 and other lipoxygenase products while PGE2 (in the presence of LTs) acts to facilitated ear swelling, although serotonin and histamine may also contribute.
Polyunsaturated alkamides isolated from Achillea species, Echinacea angustifolia DC., Anacyclus pyrethrum (L.) Link, and Aaronsohnia pubescens (Desf.) Bremer & Humphries, (Compositae) were shown to possess inhibitory activity in in vitro cyclooxygenase (sheep seminal microsomes) and 5-lipoxygenase (porcine leukocytes) assays. Activity appeared to depend on the particular structure of the alkamides.
Buddlejasaponin I and saikosaponin 1 and 2, biologically active compounds from Scrophularia scorodonia and Bupleurum rigidum respectively, exert potent in vivo antiinflammatory effects on mouse ear edema induced by phorbol myristate acetate (PMA). The effects of these compounds on swelling and other inflammatory parameters are described. In screening for in vitro effects of saikosaponins on cellular systems generating cyclooxygenase (COX) and lipoxygenase (LOX) metabolites, we observed that most saikosaponins showed a significant effect. The action is more marked on LOX metabolite LTC4. Our data support the inhibition of arachidonic acid metabolism as one of the biochemical mechanisms that might be the rationale for the putative antiphlogistic activity of these saikosaponins.
The bioactive amides affinin and capsaicin isolated respectively from Heliopsis longipes roots and Capsicum spp fruits, were assayed for activity against Escherichia coli, Pseudomonas solanacearum, Bacillus subtilis and Saccharomyces cerevisicae suspension cultures. The alkamide affinin inhibited growth of E. coli and S. cerevisiae at concentrations as low as 25 microg/ml. Higher concentrations of affinin were necessary to inhibit growth of P. solanacearum and B. subtilis. However. high concentrations of capsaicin only retarded the growth of E. coli and P. solanacearum, whereas growth of B. subtilis was strongly inhibited and that of S. cerevisiae was initially enhanced. Results are discussed in relation to previous reports concerning crude extract and to the molecular structures of the bioactive compounds.
Wirkung von Phycocyanin-Extrakt auf die Prostaglandin E2-Spiegel am Mausohrentzündungsmodell
Vor kurzem wurde gezeigt, daß Phykocyanin, ein aus Spirulina Microalgae isoliertes Gallenprotein, einen guten Effekt gegen Entzündung in mehreren Tiermodellen ausübt. In der vorliegenden Studie wird über die Phykocyanin-Wirkung auf Konzentrationen von Prostaglandin E2 (PGE2) und die Bestimmung der Aktivität von Phospholipase A2 (PLA2) beim Arachidon-säure (AA)- und 12-O-Tetradecanoyl-phorbol-13-acetat auf (TPA)-induziertem Mausohrödem berichtet. Phykocyanin (50-200 mg/kg p.o.) hemmt in dosisabhängiger Weise die PGE2-Spiegel in mit AA behandelten Mausohren. Phycocyanin (100-400 mg/kg p.o.) verringert gemäßigt die PLA2-Aktivität in TPA-induzierten Enzündungstests am Mausohr. Triamcinolon 810 mg/ kg p.o.) das als Vergleichssubstanz eingesetzt wurde, zeigte einen bemerkenswerten Hemmeffekt auf die PLA2-Aktivität. Diese Ergebnisse liefern den ersten Beweis dafür, daß die entzündungshemmenden Effekte von Phykocyanin zumindest teilweise auf der Hemmung der PGE2-Erzeugung und der PLA2-Aktivität beruhen.
This work demonstrates the fungistatic and bacteriostatic activities of affinin, the main alkamide of Heliopsis longipes (Gray) Blake (Asteraceae) roots and two alkamides obtained by catalytic reduction of affinin: N-isobutyl-2E-decenamide and N-isobutyl-decanamide. The bioactivity was tested against Rhizoctonia solani groups AG3 and AG5, Sclerotium rolfsii, Sclerotium cepivorum, Fusarium sp., Vertcillium sp., phytopathogenic fungi; Phytophthora infestans, a phytopathogenic Chromista; Saccharomyces cerevisiae, a nonphytopathogenic ascomycete; and Escherichia coli, Erwinia carotovora, and Bacillus subtilis, bacteria. Affinin, being the primary component of the lipidic fraction, is expected to be responsible for the fungitoxic activity observed in roots of this plant species. Four of the assayed fungi showed an important sensitivity to the presence of affinin: S. rolfsii, S. cepivorum, P. infestans, and R. solani AG-3 and AG-5, displaying a growth inhibition of 100%. S. cerevisiaeshowed a similar growth inhibition with affinin. None of the alkamides obtained by catalytic reduction of affinin showed a fungitoxic activity. Affinin had a definite negative effect on the growth of E. coli and B. subtilis, but E. carotovora carotovora was not sensitive to the highest dose of affinin assayed. N-Isobutyl-2E-decenamide displayed a higher bacteriostatic activity against E. coli and E. carotovora carotovora. In both cases, this alkamide was more potent than affinin. On the other hand, only N-isobutyl-decanamide displayed a significant activity on the growth of B. subtilis.
Ethnomedical questionnaires were distributed in Chicago, Costa Rica, and Colombia to identify the most common over-the-counter (OTC) plant or plant-based products advocated for treating oral pain, ulcerative conditions, and cancer within these locations. Over 100 plants or plant-based herbal preparations and commercial products, purchased from local botanical markets and pharmacies, were advocated for the treatment of oral medicine conditions. Locally familiar and common language names were attributed to the plant products at the time of purchase. Plant products or plant-based commercial products containing plant-based essential oils, anesthetic constituents, and or chemical compounds recommended as OTC oral medicine preparations were systematized, tabulated, and correlated with the published phytotherapeutic literature. Though pharmacognostic research is available for some of the species collected, further ethnographic research is needed to correlate common names with the accurate taxonomic identification for each plant species. Furthermore, epidemiological research is needed to verify the use and standardized dosage for OTC ethnomedicine preparations for oral medicine conditions. Pharmacognostic research and clinical trails which can verify taxonomy, dose, safety, active principles, and efficacy of these OTC oral medicine products must be enhanced in order to verify the claimed validity in contemporary, global, oral medicine practice.
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