ABSTRACT The herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) occur worldwide. Infections caused by these viruses have great public health importance due to the growing resistance to the first-choice drug, acyclovir, especially in immunosuppressed patients. Alkaloids derived from species of Annonaceae have been reported as antiviral agents against HSV and others viruses. Within this context, we evaluated the antiviral activity of the total alkaloid fraction (TAF) extracted from the branches of Fusaea longifolia (Aubl.) Saff. (Annonaceae), a species native to the Amazon region, against the HSV-1 and HSV-2 viruses. The antiviral activity was evaluated through the plate reduction assay and the mode of action was investigated by a set of other assays. The TAF was active against the HSV-2 strain 333 and against the HSV-1 strains KOS and 29R (acyclovir resistant), with selectivity index values (SI = 50% cytotoxic concentration/50% effective concentration) of 5, 4 and 3, respectively. In the preliminary study of the anti-HSV-2 mode of action, TAF showed viral inhibitory effects if added up to 12 h post-infection, had virucidal activity and did not present viral inhibition in pre-treatment. Our results showed that the TAF exhibited anti-HSV activity. Regarding HSV-2, TAF acted after the viral infection and had virucidal activity. A mass spectrometry analysis revealed the presence of nine alkaloids in the TAF that had previously been reported for Annonaceae, including liriodenine, lysicamine and isoboldine, which have been described as potential anti-HSV-1 agents.

RESUMO Os vírus herpes simplex tipo 1 (HSV-1) e tipo 2 (HSV-2) têm ampla ocorrência global. As infecções causadas por esses vírus têm grande importância em saúde pública devido à crescente resistência ao fármaco de primeira linha, aciclovir, principalmente em pacientes imunossuprimidos. Alcaloides derivados de espécies de Annonaceae têm sido relatados como agentes antivirais contra o HSV e outros vírus. Neste contexto, nós avaliamos a atividade antiviral da fração alcaloide total (TAF) dos ramos de Fusaea longifolia (Aubl.) Saff. (Annonaceae), uma espécie nativa da região amazônica, contra os vírus HSV-1 e HSV-2. A atividade antiviral foi avaliada através do ensaio de redução em placa e o modo de ação foi investigado por um conjunto de ensaios. O TAF foi ativo contra a cepa HSV-2 333 e contra as cepas HSV-1 KOS e 29R (resistente ao aciclovir), com valores de índice de seletividade (IS = 50% concentração citotóxica/50% concentração efetiva) de 5, 4 e 3, respectivamente. No estudo preliminar do modo de ação da atividade anti-HSV-2, o TAF inibiu a replicação viral quando adicionados até 12 h pós-infecção, apresentou atividade virucida e não apresentou inibição viral no pré-tratamento. Nossos resultados mostraram que o TAF exibiu atividade anti-HSV. Em relação ao HSV-2, o TAF atuou após a infecção viral e apresentou atividade virucida. Uma análise do TAF por espectrometria de massas identificou a presença de nove alcaloides, incluindo liriodenina, lisicamina e isoboldina, que já foram descritos como potenciais agentes anti-HSV-1.

The seed oil of Annona salzmannii A. DC. was analyzed by GC-MS and 1H qNMR, revealing a mixture of unsaturated (80.5%) and saturated (18.7%) fatty acids. Linoleic (45.3%) and oleic (33.5%) acid were the major unsaturated fatty acids identified, while palmitic acid (14.3%) was the major saturated fatty acid. The larvicidal effects of A. salzmannii seed oil were evaluated against third-instar larvae of Aedes aegypti (Linn.). The oil exhibited moderate larvicidal activity, with a LC50 of 569.77 ppm (95% CI = 408.11 to 825.88 ppm). However, when the cytotoxic effects of the oil were evaluated, no expressive antiproliferative effects were observed in tumor cell lines B16-F10 (mouse melanoma), HepG2 (human hepatocellular carcinoma), K562 (human chronic myelocytic leukemia), HL-60 (human promyelocytic leukemia), and non-tumor cell line PBMC (peripheral blood mononuclear cells), with IC50 values > 50 μg·mL-1. This is the first study to evaluate the chemical composition, larvicidal and cytotoxic activity of A. salzmannii seed oil.

The phytochemical investigation of the hexane and methanol extracts from the bark of Duguetia pycnastera Sandwith (Annonaceae) afforded seven known compounds, two asarone-derived phenylpropanoids and five isoquinoline-derived alkaloids. The asarones, γ-asarone (1-allyl-2,4,5-trimethoxybenzene) and 2,4,5-trimethoxy-styrene were isolated of the hexane extract while the aporphine alkaloids, O-methylmoschatoline, lysicamine, nornuciferidine, and guatterine N-oxide, and the benzyltetrahydroisoquinoline alkaloid, (S)-reticuline were isolated of the alkaloid fraction of the methanol extract. This is the first report of these compounds in D. pycnastera. γ-Asarone is being reported for the first time in the Annonaceae. Nornuciferidine is described for the second time in the Annonaceae while guatterine N-oxide is the third register. The structures of the isolated compounds were established by extensive analyses using 1D and 2D NMR spectroscopy in combination with MS. The cytotoxic activity of the isolated compounds (except for nornuciferidine) was evaluated against cancer and non-cancerous cell lines, in which lysicamine was the most active compound, mainly against HL-60, HepG2, and K562 with IC50 values of 24.40, 28.86 and 38.75 µmol L-1, respectively.

The phytochemical investigation of the alkaloid-rich fraction obtained from the leaves of Guatteria pogonopus Mart. (Annonaceae) allowed the isolation and identification for the first time in this species of: (+)-nornuciferine (1), a mixture of 1 and (+)-anonaine (2), (+)-isocorydine (3), (+)-nuciferine (4), (+)-roemerine (5), (-)-tetrahydropseudocolumbamine (6), a mixture of 6, liriodenine (9) and lysicamine (10), a mixture of 1,2,9-trimethoxy-10-hydroxyaporphine (7) and bulbocapnine (8), 9, 10, and (+)-N-methyllindicarpine (11). Compounds 6, 7, 8, and 11 have not been previously reported in the family Annonaceae. Furthermore, the formerly synthetic 1,2,9-trimethoxyaporfin-10-ol (7) is described for the first time as a natural aporphine alkaloid herein. The chemical structures were established by 1D and 2D NMR as well as in comparison with data previously reported in the literature. The cytotoxic activity of the alkaloids was evaluated against tumor (B16-F10, HepG2, HL-60, and K562) and non-tumor (PBMC) cell lines. Alkaloid 1 presented significant activity against HepG2 cell lines with IC50 of 9.60 µmol L-1 while the mixture of 6, 9 and 10 displayed strong cytotoxic activity against HL-60 and K562 cell lines with IC50 values of 3.41 an 8.50 µmol L-1, respectively.

The chemical composition of the essential oil and hydrolates of Campomanesia viatoris Landrum were investigated by gas chromatography/mass spectrometry (GC/MS) and a GC flame ionization detector (GC-FID). The major constituents were tasmanone (70.50, essential oil; 74.73%, hydrolate), flavesone (12.77, essential oil; 12.24%, hydrolate) and agglomerone (6.79, essential oil; 10.84%, hydrolate). Tasmonone was isolated and its structure was characterized by spectrometric analysis, specifically 1D and 2D nuclear magnetic resonance (NMR) and mass spectrometry (MS). These findings supports the quimiotaxonomic relationship with Campomanesia and Eucalyptus genera.

<p>The phytochemical investigation of the stem bark of <italic>Annona pickelii</italic> yielded four steroids (β-sitostenone, β-sitosterol, stigmasterol and campesterol), three lignans (eudesmin, magnolin and yangambin), twelve alkaloids (liriodenine, lysicamine, atherospermidine, anonaine, analobine, asimilobine, discretamine, stepholidine, coclaurine, orientaline, juziphine and stepharine), and a benzenoid (2-methoxybenzoic acid). These compounds support their recent reclassification from <italic>Rollinia</italic> to <italic>Annona</italic>, and that it is a typical species of the family Annonaceae. Significant antifungal and antioxidant activities were found for the methanol crude extract as well as for the lignans eudesmin, magnolin, yangambin and the alkaloid discretamine. In addition, several items of NMR data for the alkaloids were reviewed and unequivocally described in this work.</p>

Analysis of the volatile fraction of Aristolochia trilobata stem led to the identification of 6-methyl-5-hepten-2-yl acetate (23.31 ± 0.28%), limonene (15.43 ± 0.030%), linalool (8.70 ± 0.29%), p-cymene (7.81 ± 0.12%), bicyclogermacrene (4.21 ± 0.11%), and spathulenol (4.17 ± 0.14%) as the major constituents of the essential oil. Linalool (29.51 ± 0.49%), 6-methyl-5-hepten-2-ol (19.54 ± 0.82%), 6-methyl-5-hepten-2-yl acetate (8.92 ± 0.16%), and a-terpineol (4.62 ± 0.05%) were identified as major constituents of the hydrolate. The compound 6-methyl-5-hepten-2-yl acetate was isolated for the first time from this plant and was identified as the major component of the volatile fraction.

A investigação fitoquímica das folhas de Guatteria friesiana (Annonaceae) levou à obtenção de três novos alcaloides isoquinolínicos, 13-hidroxi-discretinina, 6,6a-desidroguateriopsiscina e 9-desidroxi-1-metoxi-diidroguattouregidina, juntamente com oito alcaloides conhecidos, 13-hidroxi-2,3,9,10-tetrametoxiprotoberberina, guateriopsiscina, lisicamina, liriodenina, aterospermidina, lanuginosina, 7,8-diidro-8-hidroxipalmatina e palmatina. 13-Hidroxi-2,3,9,10-tetrametoxiprotoberberina, obtido somente através de síntese, é relatada pela primeira vez como produto natural. As estruturas dos alcaloides isolados foram elucidadas por extensivas análises de ressonância magnética nuclear (RMN 1D e 2D), espectrometria de massas (MS) e comparação com os dados descritos na literatura. A atividade citotóxica in vitro dos alcaloides majoritários foi avaliada frente a linhagens de células tumorais e não tumorais. Considerando a atividade média, de acordo com os critérios do National Cancer Institute (NCI/EUA), todos os alcaloides avaliados foram inativos. Entretanto, o alcaloide palmatina apresentou efeito citostático para as linhagens MCF-7 (mama) e U251 (glioma) com valores de GI50 abaixo de 20,0 µmol L-1 (10,5 e 16,2 µmol L-1, respectivamente), sugerindo uma ação citotóxica seletiva.

Phytochemical investigation of the leaves of Guatteria friesiana (Annonaceae) afforded three new isoquinoline alkaloids, 13-hydroxy-discretinine, 6,6a-dehydroguatteriopsiscine and 9-dehydroxy-1-methoxy-dihydroguattouregidine. Eight known alkaloids were also isolated, 13-hydroxy-2,3,9,10-tetramethoxyprotoberberine, guatteriopsiscine, lysicamine, liriodenine, atherospermidine, lanuginosine, 7,8-dihydro-8-hydroxypalmatine and palmatine. 13-Hydroxy-2,3,9,10-tetramethoxyprotoberberine was only obtained by synthesis and is being reported as a natural product for the first time. The structures of the isolated alkaloids were established by extensive analysis of 1D and 2D nuclear magnetic resonance (NMR) and mass spectrometric (MS) data, as well as by comparison with data reported in the literature. The in vitro cytotoxic activity of the major alkaloids was evaluated against tumor and non-tumor cell lines. All of the alkaloids evaluated were determined to be inactive based on National Cancer Institute (NCI/USA) criteria. However, the alkaloid palmatine exhibited a cytostatic effect on MCF-7 (breast) and U251 (glioma) human tumor cell lines, with GI50 values lower than 20.0 µmol L-1 (10.5 and 16.2 µmol L-1, respectively), suggesting a selective cytotoxic action.

The dereplication of aporphine and oxoaporphine alkaloids by direct infusion in ESI-IT-MSn system was applied for alkaloidal fractions of the Unonopsis guatterioides (Annonaceae). Its main advantage over other dereplication methods is the ability to quickly identify substances in complex mixtures without the use of coupled techniques and expensive databases. By only the fragmentation keys and comparison with literature data the aporphine alkaloids anonaine, asimilobine and nornuciferine were identified. The nornuciferine is being reported for the first time in Unonopsis. The oxoaporphine alkaloids liriodenine and lisycamine were identified in the alkaloidal fractions by comparison with the fragmentations of authentic samples.

The biological activity of seven extracts from leaves of different Jatropha curcas L (Euphorbiaceae) accessions was evaluated on Spodoptera frugiperda. Methanol extracts were incorporated into an artificial diet and offered to the larval stage of S. frugiperda. The parameters evaluated were length of larval and pupal stages, mortality of larval and total cycle stage, and weight of pupae. The extracts of the EMB accessions showed the best result for larval mortality at 60.00 and 56.67%, compared with the control, respectively. Hexane partition of the methanol extract of the leaves of PM-14 accessions allowed the identification of phytosterols, phytol and n-alkanols.

Phytochemical investigation of the hexane extract from the stem of Xylopia laevigata led to the isolation of the ent-kaurane diterpenoids, ent-kaur-16-en-19-oic acid, 4-epi-kaurenic acid, ent-16β-hydroxy-17-acetoxy-kauran-19-al, ent-3β-hydroxy-kaur-16-en-19-oic acid, and ent-16β,17-dihydroxy-kauran-19-oic acid, as well as spathulenol and a mixture of β-sitosterol, stigmasterol and campesterol. The identification of the compounds was performed on the basis of spectrometric methods including GC-MS, IR, and 1D and 2D NMR. Potent larvicidal activity against Aedes aegypti larvae with LC50 of 62.7 µg mL-1 was found for ent-3β-hydroxy-kaur-16-en-19-oic acid. This compound also showed significant antifungal activity against Candida glabrata and Candida dubliniensis with MIC values of 62.5 µg mL-1.

bicyclogermacrene (35.12%), (E)-caryophyllene (14.19%) and α-copaene (8.19%). The antimicrobial and antileishmanial activities were investigated. The oil showed potent antimicrobial activity against Candida albicans and Rhodococcus equi. The oil also showed significant antileishmanial activity, giving the best results against Leishmania guyanensis. A preliminary cytotoxicity assay for this oil was carried out on hamster and mice (Balb/c) peritoneal macrophages. The results obtained were similar to pentamidine and considered not to be cytotoxic to macrophages.