In Vitro Leishmanicidal Activity of Copaiba Oil and Kojic Acid Combination on the Protozoan Leishmania (Leishmania) amazonensis and Host Cell.

(1) Background: Leishmaniasis refers to a group of anthropozoonotic diseases caused by Leishmania. The major chemotherapeutic agent used for its treatment is Glucantime®®, but the search continues for new compounds that are economically viable and act on the protozoan without causing damage to the host cell. As an alternative approach, this study used a combination of copaiba oil (CO) and kojic acid (KA) to determine their in vitro action on host cells, on the Leishmania (Leishmania) amazonensis protozoan and its interaction with macrophages. (2) Methods: In vitro culture, analysis of cytokine release and microscopy assays were performed. Statistical analysis was performed with ANOVA (GraphPad Prism). (3) Results: The combination did not induce cytotoxic effects on macrophages after treatment but promoted morphological changes in the protozoan, such as nuclear alterations (apoptotic characteristics), alterations in the cellular body and an increase in the number of electrodense structures and acidocalcisomes, observed mainly at the concentrations of CO20KA50 and CO30KA50 µg/mL. We observed reductions in the intracellular amastigote number and in the production of proinflammatory cytokines, such as IL-6 and TNF-α, after treatment with CO30KA at 50 µg/mL. (4) Conclusions: We report here, for the first time, that the combination of CO and KA may be a promising approach against Leishmania (Leishmania) amazonensis.

Exploring the chemical composition, in vitro and in silico study of the anticandidal properties of annonaceae species essential oils from the Amazon.

Chemical composition of the essential oils (EOs) from the leaves of five Annonaceae species found in the amazon region was analyzed by Gas chromatography coupled to mass spectrometry. The antifungal activity of theses EOs was tested against Candida albicans, Candida auris, Candida famata, Candida krusei and Candida tropicalis. In addition, an in silico study of the molecular interactions was performed using molecular modeling approaches. Spathulenol (29.88%), α-pinene (15.73%), germacra-4(15),5,10(14)-trien-1-α-ol (6.65%), and caryophylene oxide (6.28%) where the major constitents from the EO of Anaxagorea dolichocarpa. The EO of Duguetia echinophora was characterized by ß-phellanderene (24.55%), cryptone (12.43%), spathulenol (12.30%), and sabinene (7.54%). The major compounds of the EO of Guatteria scandens where ß-pinene (46.71%), α-pinene (9.14%), bicyclogermacrene (9.33%), and E-caryophyllene (8.98%). The EO of Xylopia frutescens was characterized by α-pinene (40.12%) and ß-pinene (36.46%). Spathulenol (13.8%), allo-aromadendrene epoxide (8.99%), thujopsan-2-α-ol (7.74%), and muurola-4,10(14)-dien-1-ß-ol (7.14%) were the main chemical constituents reported in Xylopia emarginata EO. All EOs were active against the strains tested and the lowest inhibitory concentrations were observed for the EOs of D. echinophora, X. emarginata, and X. frutescens against C. famata the Minimum Inhibitory Concentration values of 0.07, 0.019 and 0.62 µL.mL-1, respectively. The fungicidal action was based on results of minimum fungicidal concentration and showed that the EOs showed fungicide activity against C. tropicalis (2.5 µL.mL-1), C. krusei (2.5 µL.mL-1) and C. auris (5 µL.mL-1), respectively. The computer simulation results indicated that the major compounds of the EOs can interact with molecular targets of Candida spp.

Phytochemical Profile, Antioxidant Potential and Toxicity Evaluation of the Essential Oils from Duguetia and Xylopia Species (Annonaceae) from the Brazilian Amazon.

The essential oils (EOs) of Duguetia echinophora, D. riparia, Xylopia emarginata and X. frutescens (Annonaceae) were obtained by hydrodistillation and the chemical composition was analyzed by GC-MS. An antioxidant assay using the ABTS and DPPH radicals scavenging method and cytotoxic assays against Artemia salina were also performed. We evaluated the interaction of the major compounds of the most toxic EO (X. emarginata) with the binding pocket of the enzyme Acetylcholinesterase, a molecular target related to toxicity in models of Artemia salina. The chemical composition of the EO of D. echinophora was characterized by ß-phellandrene (39.12%), sabinene (17.08%) and terpinolene (11.17%). Spathulenol (22.22%), caryophyllene oxide (12.21%), humulene epoxide II (11.86%) and allo-aromadendrene epoxide (10.20%) were the major constituents of the EO from D. riparia. Spathulenol (5.65%) and caryophyllene oxide (5.63%) were the major compounds of the EO from X. emarginata. The EO of X. frutescens was characterized by α-pinene (20.84%) and byciclogermacrene (7.85%). The results of the radical scavenger DPPH assays ranged from 15.87 to 69.38% and the highest percentage of inhibition was observed for the EO of X. emarginata, while for ABTS radical scavenging, the antioxidant capacity of EOs varied from 14.61 to 63.67%, and the highest percentage of inhibition was observed for the EO of X. frutescens. The EOs obtained from D. echinophora, X. emarginata and X. frutescens showed high toxicity, while the EO of D. riparia was non-toxic. Because the EO of X. emarginata is the most toxic, we evaluated how its major constituents were able to interact with the Acetylcholinesterase enzyme. The docking results show that the compounds are able to bind to the binding pocket through non-covalent interactions with the residues of the binding pocket. The species X. emarginata and X. frutescens are the most promising sources of antioxidant compounds; in addition, the results obtained for preliminary cytotoxicity of the EOs of these species may also indicate a potential biological activity.

Flavanone Glycosides, Triterpenes, Volatile Compounds and Antimicrobial Activity of Miconia minutiflora (Bonpl.) DC. (Melastomataceae).

Chemical composition of the essential oils and extracts and the antimicrobial activity of Miconia minutiflora were investigated. The flavanone glycosides, pinocembroside and pinocembrin-7-O-[4″,6″-HHDP]-ß-D-glucose, were identified, along with other compounds that belong mainly to the triterpene class, besides the phenolics, gallic acid and methyl gallate. Sesquiterpenes and monoterpenes were the major compounds identified from the essential oils. Screening for antimicrobial activity from the methanolic extract of the leaves showed that the MIC and MMC values against the tested microorganisms ranged from 0.625 to 5 mg·mL−1 and that the extract was active against microorganisms, Staphyloccocus aureus, Escherichia coli, and Bacillus cereus.

Essential Oils from Annonaceae Species from Brazil: A Systematic Review of Their Phytochemistry, and Biological Activities.

The present work involves a systematic review of the chemical composition and biological effects of essential oils from the Annonaceae species collected in Brazil from 2011 to 2021. Annonaceae is one of the most important botanical families in Brazil, as some species have economic value in the market as local and international fruit. In addition, the species have useful applications in several areas-for instance, as raw materials for use in cosmetics and perfumery and as medicinal plants. In folk medicine, species such as Annona glabra L. and Xylopia sericea A. St.-Hil. are used to treat diseases such as rheumatism and malaria. The species of Annonaceae are an important source of essential oils and are rich in compounds belonging to the classes of mono and sesquiterpenes; of these compounds, α-pinene, ß-pinene, limonene, (E)-caryophyllene, bicyclogermacrene, caryophyllene oxide, germacrene D, spathulenol, and ß-elemene are the most abundant. The antimicrobial, anti-inflammatory, antileishmania, antioxidant, antiproliferative, cytotoxic, larvicidal, trypanocidal, and antimalarial activities of essential oils from the Annonaceae species in Brazil have been described in previous research, with the most studies on this topic being related to their antiproliferative or cytotoxic activities. In some studies, it was observed that the biological activity reported for these essential oils was superior to that of drugs available on the market, as is the case of the essential oil of the species Guatteria punctata (Aubl.) R. A. Howard., which showed a trypanocidal effect that was 34 times stronger than that of the reference drug benznidazol.

Chemical Composition of Volatile Compounds in Apis mellifera Propolis from the Northeast Region of Pará State, Brazil.

Propolis is a balsamic product obtained from vegetable resins by exotic Africanized bees Apis mellifera L., transported and processed by them, originating from the activity that explores and maintains these individuals. Because of its vegetable and natural origins, propolis is a complex mixture of different compound classes; among them are the volatile compounds present in the aroma. In this sense, in the present study we evaluated the volatile fraction of propolis present in the aroma obtained by distillation and simultaneous extraction, and its chemical composition was determined using coupled gas chromatography, mass spectrometry, and flame ionization detection. The majority of compounds were sesquiterpene and hydrocarbons, comprising 8.2-22.19% α-copaene and 6.2-21.7% ß-caryophyllene, with additional compounds identified in greater concentrations. Multivariate analysis showed that samples collected from one region may have different chemical compositions, which may be related to the location of the resin's production. This may be related to other bee products.

Betulinic Acid Prevents the Acquisition of Ciprofloxacin-Mediated Mutagenesis in Staphylococcus aureus.

The occurrence of damage on bacterial DNA (mediated by antibiotics, for example) is intimately associated with the activation of the SOS system. This pathway is related to the development of mutations that might result in the acquisition and spread of resistance and virulence factors. The inhibition of the SOS response has been highlighted as an emerging resource, in order to reduce the emergence of drug resistance and tolerance. Herein, we evaluated the ability of betulinic acid (BA), a plant-derived triterpenoid, to reduce the activation of the SOS response and its associated phenotypic alterations, induced by ciprofloxacin in Staphylococcus aureus. BA did not show antimicrobial activity against S. aureus (MIC > 5000 µg/mL), however, it (at 100 and 200 µg/mL) was able to reduce the expression of recA induced by ciprofloxacin. This effect was accompanied by an enhancement of the ciprofloxacin antimicrobial action and reduction of S. aureus cell volume (as seen by flow cytometry and fluorescence microscopy). BA could also increase the hyperpolarization of the S. aureus membrane, related to the ciprofloxacin action. Furthermore, BA inhibited the progress of tolerance and the mutagenesis induced by this drug. Taken together, these findings indicate that the betulinic acid is a promising lead molecule in the development helper drugs. These compounds may be able to reduce the S. aureus mutagenicity associated with antibiotic therapies.

In vitro acaricidal activity of Crescentia cujete L. fruit pulp against Rhipicephalus microplus.

Cattle tick (Rhipicephalus microplus) is an important constraint on livestock production, particularly in tropical and subtropical areas. Synthetic acaricides are the primary method of control this thick. Resistance of R. microplus to synthetic acaricides has given rise to the need of new scientific investigations on alternative ways to control it. The objective of this work was to conduct a chemical investigation of Crescentia cujete L. fruit pulp and to evaluate the acaricide effect on R. microplus larvae of extracts, fractions, and isolated substances from this species. Crescentia cujete is a Bignoniaceae and it is characteristic of tropical climates. Its most distinctive feature is its fruit, which is spherical and gourd-like, with a hard woody shell and gelatinous pulp. The fruit pulp of C. cujete was extracted with ethanol by maceration yielding an extract and a residue. The extract was partitioned giving an ethyl acetate phase (EAF) that was fractionated by chromatographic procedures yielding cinnamic acid and benzoic acid. The remained botanic material from the ethanol extraction was further and successively extracted with ethyl ether and methanol under reflux. The acaricide effect of all extracts and fractions was evaluated against R. microplus using the larval packet test. The best result was observed for the EAF that caused 100% of mortality when a 10% solution was used and with LC50 of 5.9%. Cinnamic acid was also tested showing 63.0% of mortality with LC50 of 6.6% at the same concentrations and time of the experiment. The data obtained in this study provides information related to the chemical nature and the acaricide activity of the C. cujete epicarp that has not been previously reported. This study shows that the ethyl acetate phase of the ethanol extract of C. cujete fruit pulp is a potential alternative control for R. microplus and its activity is in part associated to cinnamic acid, its major compound.

Constituents and Pharmacological Activities of Myrcia (Myrtaceae): A Review of an Aromatic and Medicinal Group of Plants.

Myrcia is one of the largest genera of the economically important family Myrtaceae. Some of the species are used in folk medicine, such as a group known as "pedra-hume-caá" or "pedra-ume-caá" or "insulina vegetal" (insulin plant) that it is used for the treatment of diabetes. The species are an important source of essential oils, and most of the chemical studies on Myrcia describe the chemical composition of the essential oils, in which mono- and sesquiterpenes are predominant. The non-volatile compounds isolated from Myrcia are usually flavonoids, tannins, acetophenone derivatives and triterpenes. Anti-inflammatory, antinociceptive, antioxidant, antimicrobial activities have been described to Myrcia essential oils, while hypoglycemic, anti-hemorrhagic and antioxidant activities were attributed to the extracts. Flavonoid glucosides and acetophenone derivatives showed aldose reductase and α-glucosidase inhibition, and could explain the traditional use of Myrcia species to treat diabetes. Antimicrobial and anti-inflammatory are some of the activities observed for other isolated compounds from Myrcia.

Chemical investigation of Cyperus distans L. and inhibitory activity of scabequinone in seed germination and seedling growth bioassays.

Chemical investigation of the rhizomes of Cyperus distans (Cyperaceae) led to the identification of α-ciperone, cyperotundone and scabequinone, besides other common constituents. Complete assignment of the (13)C NMR data of scabequinone is being published for the first time. The inhibitory effects of C. distans extracts and scabequinone on the seed germination and seedling growth of Mimosa pudica, Senna obtusifolia and Pueraria phaseoloides were evaluated. Seed germination inhibition bioassay revealed that S. obtusifolia (52-53%) was more sensitive to the hexane and the methanol extracts at 1% than M. pudica (0-10%). Scabequinone at 250 mg L⁻¹ displayed seed germination inhibitions more than 50% and radicle growth reduction of more than 35% of the test species S. obtusifolia and P. phaseoloides, while the hypocotyl growth of M. pudica was significantly affected (>50%) by the quinone at the same concentration. These results demonstrate that scabequinone contributes to the overall inhibitory activities of C. distans.

Ricinine and other constituents of Aparisthmium cordatum (Euphorbiaceae).

The chemical study of Aparisthmium cordatum (Euphorbiaceae) led to the isolation of tannins, together with the alkaloid ricinine and other common compounds. The composition of A. cordatum is similar to most of the Alchornea species, from the same subtribe, except for the occurrence of ricinine. This study rectifies the first investigations published for A. cordatum that were conducted with Croton palanostigma.

Antibacterial activity of alkaloids produced by endophytic fungus Aspergillus sp. EJC08 isolated from medical plant Bauhinia guianensis.

Bauhinia guianensis is a typical plant in the Amazon region belonging to the family Leguminosea, used by local populations for the treatment of infectious and renal diseases. Previous work on the plant B. guianensis led to the isolation of substances with anti-inflammatory and analgesic activities. Thus, compounds isolated from B. guianensis with antimicrobial activities had not been identified. Given that there is a possibility of biological activity reported for a given plant being found in the endophytic fungi, we decided to isolate endophytic fungi from B. guianensis and test their antimicrobial activities. The alkaloids known as fumigaclavine C and pseurotin A were isolated by column chromatography and identified by 1D and 2D NMR techniques and mass spectrometry. The alkaloids are first reported as broad-spectrum antibacterial agents with good activity.

The genus Mansoa (Bignoniaceae): a source of organosulfur compounds / O gênero Mansoa (Bignoniaceae): uma fonte de compostos organosulfurados

The genus Mansoa belongs to the family Bignoniaceae and it includes eleven species that occur mainly in the dry and wet forests of Brazil and from Argentina to the Southeast of Mexico. These species in the Brazilian Amazon region are known as "cipó-de-alho", that means garlic vine, in reference to the pungent garlic-like smell of the leaves when crushed. "Cipó-de-alho" has several uses in folk medicine and among them, the most cited are the treatment for cold, fever, pain and inflammation of arthritis and rheumatism. In spite of all those uses, it still has little application in phytotherapy when compared to garlic (Allium sativum). The essential oils of Mansoa spp. show the presence of allyl polysufides that contribute to the characteristic aroma and flavor. The chemical composition of the organic extracts of Mansoa has been reported and it includes alkanes, alkanols, triterpenoids, flavonoids, lapachol derivatives and organosulfur compound alliin. The uses, chemical composition, biological activities and agricultural aspects of Mansoa species and their relationship with A. sativum are presented.

O gênero Mansoa pertence à família Bignoniaceae e inclui onze espécies que ocorrem principalmente nas florestas secas e úmidas do Brasil e da Argentina até o Sudeste do México. Essas espécies na Amazônia brasileira são conhecidas como "cipó-de-alho", em referência ao forte cheiro de alho das folhas quando esmagadas. O "cipó-de-alho" tem vários usos na medicina tradicional e entre eles, os mais citados são para tratamento de gripe, febre, dor e inflamação de artrite e reumatismo. Apesar de todos os usos, ainda tem pequena aplicação como fitoterápico quando comparado ao alho (Allium sativum). Os óleos essenciais de Mansoa spp. contêm polissulfetos de alila que contribuem para o aroma e sabor característicos. A composição química dos extratos orgânicos de Mansoa incluiu alcanos, alcanóis, triterpenóides, flavonóides, derivados do lapachol e o derivado sulfurado aliína. Os usos, composição química, atividades biológicas e aspectos agrícolas de espécies de Mansoa e sua relação com A. sativum são apresentados.