BACKGROUND:Malaria is a deadly parasitic disease for which innovative treatments are urgently needed. A mixture of eight triterpenic esters (8TTE) was previously identified as important for the antiplasmodial activity of Keetia leucantha twigs, a plant used in traditional medicine in Benin. Despite the reported in vitro and in vivo activity, the targets of 8TTE are unknown. METHODS:The present study investigated the mode of action of 8TTE on Plasmodium falciparum by a multi-scale integrative study from phenotype to metabolome, including: phenotypic analysis, enzymatic tests, molecular docking and metabolomic profiling. RESULTS:This study identified a unique antiplasmodial profile with activity onset in the early-ring stage of the parasite, the inhibition of aminopeptidase PfA-M17 (PlasmoDB PF3D7_1446200) and perturbations in parasite haemoglobin metabolism. CONCLUSIONS:Further structure-activity and medicinal chemistry studies are warranted to elaborate on these findings and the potential for 8TTE-related molecules to serve as future antimalarial drugs.

The genus Psiadia (Asteraceae), widely distributed in Madagascar and the Mascarene Islands (Mauritius, La Réunion, Rodrigues), is traditionally used to treat bronchitis, asthma, colds, abdominal pain, and other inflammatory disorders. However, few studies have scientifically validated these traditional medicinal uses. To assess P. dentata as a valuable source of bioactive natural products, a combined 1H NMR-based metabolomic, molecular networking, and phytochemical study was conducted. Multivariate analysis (PLS-DA) of crude extracts from Psiadia species collected on Reunion Island enabled rapid discrimination of active extracts from P. dentata and revealed two methoxylated flavonoids and one coumarin as metabolites correlated with its antiplasmodial and anti-inflammatory activities. Additionally, UHPLC-DAD-ESI-QTOF-MS/MS molecular networking approach enabled detailed chemical profiling of this species, allowing the annotation of 25 compounds (1–25) in this species. Subsequent phytochemical investigation of P. dentata leaves led to the isolation and identification of 25 metabolites, including nine new diterpenes (26–34), one new coumarin (35), and 15 known compounds (1–8, 11, 18, 19 and 36–39) from the diterpenoid, flavonoid, and coumarin families. The structures of the new compounds were elucidated using spectroscopic methods, including extensive 1D and 2D NMR and HRESIMS analyses. Biological evaluation of the isolated compounds showed that compounds 1, 7, 26 and 27 showed antiplasmodial activity against Plasmodium falciparum (3D7 strain, IC50 = 7.25–13.46 μM). Compounds 7, 26, 27, 31 and 32 inhibited nitric oxide production (IC50 = 0.87–27.71 μM), indicating potential anti-inflammatory effects. Only compound 1 displayed moderate cytotoxicity against HepG2 and HT29 cancer cell lines (IC50 = 25.67 and 18.35 μM, respectively).

Abstract Background Triple-negative breast cancer (TNBC) presents significant therapeutic limitations due to its aggressive heterogeneity and the rapid emergence of adaptive resistance to apoptosis-based regimens. Addressing these challenges requires polypharmacological strategies capable of modulating multiple signalling networks simultaneously. While the Cannabis sativa phytocomplex offers a vast chemical space for multi-target intervention, the quantitative pharmacological basis of its synergistic interactions remains largely uncharacterised. Purpose This study aimed to deconstruct the synergistic landscape of high-purity phytocannabinoids (CBD, CBG, CBD-A) in combination with the sesquiterpene β-caryophyllene (BCP) against TNBC, using MDA-MB-231 as a primary model and Hs578T as a validation line. Methods Growth Rate (GR) inhibition metrics and the SynergyFinder+ framework were used to map pharmacological interactions across four reference models. Subcellular dynamics and phenotypic transitions were characterised by high-resolution label-free holotomographic microscopy combined with live-cell kinetic imaging and single-cell fate mapping. Results Two highly potent synergistic clusters were identified for CBD-CBG-BCP combinations, with ZIP, HSA, and Bliss synergy scores exceeding 65. CBD-A exhibited minimal interaction potential and was excluded from ternary studies. GR-based quantification further revealed that these combinations produced net cytotoxicity (GR < 0) at sub-IC₅₀ concentrations of each component. Single-cell fate mapping by holotomographic microscopy identified a temporally ordered death programme: an initial phase of extensive cytoplasmic vacuolisation associated with focal perinuclear space swelling and progressive nuclear compression, morphological hallmarks of autosis, which is followed by a transition to apoptotic execution. The autotic nature of the primary death phase was confirmed by pharmacological rescue with digoxin, a selective inhibitor of the Na⁺,K⁺-ATPase. To the best of our knowledge, this sequential engagement of autosis followed by apoptotic execution represents the first documented instance of such a two-stage death programme in any cellular model. Conclusion These findings provide robust evidence that specific phytocannabinoid-terpene ratios engage a Na⁺,K⁺-ATPase-regulated autotic programme as an upstream commitment step, followed by apoptotic execution, effectively circumventing the caspase-independent resistance mechanisms characteristic of TNBC. This study establishes a rational, quantitatively validated framework for transitioning from empirical botanical use to evidence-based, multi-target cannabinoid polypharmacology in aggressive breast cancer.

IntroductionUrsolic acid (UA), the antimalarial triterpenic mixture 8TTE (containing C-27 feruloyl and coumaroyl esters of ursane and oleane skeletons), and the semi-synthetic antitrypanosomal derivative ursolic acid O-phenyl propionate (UAOPP) exhibit high lipophilicity, which may limit their oral bioavailability. This study aimed to develop lipid nanocapsules (LNCs) to improve the solubility, intestinal permeability, and antiparasitic activity of these triterpenic compounds for potential oral delivery.MethodsLNCs formulations containing UA, 8TTE, and UAOPP were prepared and evaluated. A sensitive UPLC\x{2013}MS method was developed and validated for selected triterpenes quantification during transport studies across Caco-2 cell monolayers [limit of detection (LOD): 2 nM; limit of quantification (LOQ): 25 nM]. Cytotoxicity and permeability studies were conducted on Caco-2 cells to assess formulation safety and intestinal transport. In vitro antiparasitic activity of free and formulated compounds was evaluated against Plasmodium falciparum and Trypanosoma brucei brucei (Tbb).Results and discussionThe formulations were non-toxic to Caco-2 cells at concentrations up to 2 mg/mL. Permeability studies demonstrated enhanced transport for the formulated triterpenic esters, with permeability increases of up to 2.68-fold, shifting their classification from poorly absorbed to moderately absorbed compounds in humans [apparent permeability coefficient (Papp) > 1 × 10–6 cm/s]. Free UA showed the highest Papp value (4.95 × 10–6 cm/s ± 1.29 × 10–7), but caused epithelial integrity disruption after 2 h of incubation and during the following 48 h, whereas formulated UA induced minimal integrity loss at the same concentration. In antiparasitic assays, blank LNCs exhibited maximum non-toxic concentrations of 165 μg/mL against P. falciparum and 65 μg/mL against Tbb. At these maximum concentrations, formulated UA and 8TTE showed enhanced antiplasmodial activity; however, blank LNCs produced comparable effects. In contrast, UAOPP-loaded LNCs showed significantly improved antitrypanosomal activity by approximately 20% at 2.15 μM (cell viability: 38.20% ± 5.41) compared with free UAOPP (20.38% ± 8.80). These findings suggest that LNCs represent promising oral delivery systems for lipophilic triterpenes. Further in vivo pharmacokinetic and efficacy studies are needed to confirm their therapeutic potential.

Despite ongoing efforts, malaria persists as a significant health challenge in developing countries. This study aims to investigate the antimalarial potential of Alstonia boonei stem bark, a traditional plant in Cameroon. The EtOH-H2O (70:30) extract was subjected to phytochemical screening and chromatographic isolation of compounds. In vitro antiplasmodial activity was assessed in culture on Plasmodium falciparum using the lactate dehydrogenase assay. Colorimetric resazurin-based assay was used to assess the selectivity on LLC-MK2 monkey kidney epithelial cells. The acute oral toxicity of the crude extract was assessed in animals, as well as the in vivo antimalarial potency in Plasmodium berghei-infected mice. Two novel alkaloids, picralimine (1) and picralimine acetate (2), were isolated along with betulinic acid, sitosterol 3-O-beta-D-glucopyranoside, and a mixture of stigmasterol and sitosterol. Compared to the reference drug (artemisinin; IC(50)PfW2 = 0.12 +/- 0.05; IC(50)PfCAM06 = 0.05 +/- 0.00; IC(50)PfIPC = 0.06 +/- 0.01 & micro;g/mL), which strongly inhibited the growth of each Plasmodium falciparum species, the crude extract demonstrated significant in vitro antiplasmodial activity against P. falciparum W2mef and IPC strains (IC50: 6.89 +/- 2.66 and 7.22 +/- 3.01 & micro;g/mL, respectively) with a selectivity index of 3.64. Compound 1 showed potent activity (IC50: 3.23 +/- 0.18, 4.00 +/- 4.12, and 5.38 +/- 1.91 & micro;g/mL against PfIPC, PfCAM06, and PfW2, respectively) and was noncytotoxic (SI > 500). Compound 2 exhibited weak antiplasmodial activity (IC50 6.05 +/- 3.03, 11.45 +/- 3.95, and 10.33 +/- 4.70 & micro;g/mL, respectively, against PfW2, PfIPC, and PfCAM06). The extract was nontoxic (LD50 > 2000 mg/kg) and exhibited dose-dependent suppressive and prophylactic antimalarial effects in P. berghei-infected mice, with inhibition percentages of 94.21%, 93.42%, and 96.38% at all the doses (100, 200, and 400 mg/kg), respectively. The prophylactic antimalarial test of the extract in infected rats exhibited inhibition of the parasite growth with inhibition percentages of 89.65%, 98.52%, and 98.52% at all the doses, respectively. These results demonstrate the efficacy of A. boonei extract, suggesting its potential for antimalarial phytodrug development.

Emerging RNA viruses continue to pose major global health threats, underscoring the urgent need for broad-spectrum antiviral agents. Usambarensine, a bis-indole alkaloid isolated from Strychnos usambarensis, has historically been investigated for its antimalarial and antihelminthic properties, but its antiviral potential has remained unexplored. In this study, usambarensine was isolated through bioguided fractionation and structurally confirmed by HPTLC, HPLC-UV, FT-IR, MS/MS, and NMR. The compound was first evaluated against representatives of three major RNA virus families: Coronaviridae (SARS-CoV-2), Flaviviridae (West Nile virus), and Orthomyxoviridae (Influenza A) using Vero E6 cells. Antiviral activity was quantified by RT-qPCR at 24, 48, and 72 h postinfection. Usambarensine showed strong, dose-dependent inhibition of SARS-CoV-2, achieving ∼4log10 reduction at 72 h, transient inhibition of West Nile virus and no significant activity against Influenza A. Based on the robust activity observed against SARS-CoV-2, screening was expanded within the Coronaviridae family to include SARS-CoV-1 and MERS-CoV, both of which yielded similar results. This selective antiviral profile, as well as the observation that viral RNA levels remain similar to the initial viral inoculum, suggests that usambarensine interferes with a coronavirus-specific early entry or fusion process, potentially involving a shared entry pathway in Vero E6 cells. These findings identify usambarensine as a promising natural product with selective antiviral activity against coronaviruses. Further mechanistic studies, including determining the specific stage of the viral replication cycle affected, elucidating the molecular target, and performing structure-activity relationship optimization, are needed to assess its potential as a lead scaffold for coronavirus entry inhibitors.

The aim of the present study was to isolate and characterize compounds responsible of the antiradical and antibacterial activities in the hydro-ethanolic crude extract of Erythrina excelsa stem bark. Chemical investigation of this polar extract resulted in the isolation of a fructose derivative identified as ethyl-β-D-fructofuranoside (3) and an alkaloid designated L-(+)-Abrine (4); two compounds isolated here for the first time from the genus Erythrina, along with a mixture of sitosterol 3-O-β-D-glucopyranoside (2) and stigmasterol 3-O-β-D-glucopyranoside (1). Their structures were determined using spectroscopic methods (IR, 1 2D NMR) as well as HR-ESI-MS and literature data. The hydro-ethanolic extract and compound 3 were tested for antibacterial activity on Escherichia coli, Salmonella typhi, Salmonella typhimurium and Salmonella enterica, and for antioxidant activity using free radical scavenging (DPPH) and Ferric Reducing Ability Power (FRAP). Compound 3 showed good antiradical potential (IC50 21.11 ± 0.53 μg/mL) and moderate antibacterial activity against all the tested bacteria with MIC of 200 μg/mL. For the FRAP method, hydro-ethanolic extract showed weak activity with value of 425 mg GAE/g, while Ethyl-β-D-fructofuranoside (3) exhibited a very good reducing power (40 mg GAE/g). Ethyl-β-D-fructofuranoside (3) could, therefore, be responsible for the antiradical activity of the hydro-ethanolic crude extract.

A previously undescribed iboga-indole alkaloid, voacamine A (1), and the known alkaloids, voacamine (2), voacorine (3), and voacangine (4) were isolated from the stem bark of Voacanga africana. The structures of the alkaloids were established by one- and two-dimensional nuclear magnetic resonance analyses as well as by comparison with published data. The antiplasmodial activity of the alkaloids was assessed on a chloroquine-sensitive 3D7 Plasmodium falciparum (Pf3D7) strain, and the half-maximal inhibitory concentration (IC50) values were determined. Alkaloids 2 and 3 showed good in vitro antiplasmodial activity with IC50 values of 5.734 ± 1.365 and 5.319 ± 2.206 µg/mL, respectively, at the concentration tested. A plausible biogenetic pathway of the bisindole alkaloids has been proposed. Protein-ligand co-folding was used to generate protein-ligand complexes for the pfATP6 protein target and the isolated alkaloids. The stability of the modelled complexes was evaluated using molecular dynamics simulation. Scoring metrics of the generated complexes coupled to molecular mechanics generalized-born solvation rescoring of the modelled ligand poses were employed to explain the antimalarial structure-activity relationship for the investigated compounds. Alkaloids 2 and 3 can be considered as possible scaffolds for designing new antiplasmodial lead compounds.

ETHNOPHARMACOLOGICAL RELEVANCE:Erigeron bonariensis L. (anc. Conyza bonariensis (L.) Cronquist) is a medicinal plant with traditional use in Cameroonian folk medicine and other parts of Africa for the management of ailments, including malaria. AIM:This study focused on investigating the antiplasmodial, gametocytocidal properties and toxicity profile of E. bonariensis. MATERIALS AND METHODS:Extracts of E. bonariensis were prepared by sequential maceration using n-hexane, ethyl acetate, and methanol, and their antiplasmodial properties were assessed on both sexual and asexual Plasmodium parasite stages. Additionally, cytotoxicity was assessed on the MDA-MB-231 breast cancer cell line, while acute and sub-chronic toxicity were evaluated using albino mice. RESULTS:Generally, all extracts were active against both stages of the parasites at different levels. Hexane extract exhibited good antiplasmodial activity (IC50 6.02 ± 1.13 μg/mL) against asexual parasites while also demonstrating the highest gametocytocidal activity (IC50 1.863 ± 0.717 μg/mL). All extracts were relatively non-toxic on the MDA-MB-231 breast cancer cell line. No significant adverse effects at the tested dose were observed following an acute exposure with an LD50 > 5000 mg/kg BW. In the sub-chronic toxicity evaluation, no significant changes in body weight, or hematological and biochemical parameters were observed across the dosed groups compared to the control, with a p-value <0.05. Additionally, histopathological examination of the liver indicated no signs of toxicity or abnormalities. CONCLUSION:The findings show that E. bonariensis has effective antiplasmodial and gametocytocidal properties in vitro and has relatively low toxicity. This study supports the use of E. bonariensis as a culturally appropriate alternative for malaria management. However, further studies are needed on the safety profile and in harnessing the therapeutic potential of E. bonariensis for the development of novel antimalarials.

Sickle cell disease (SCD) is a neglected tropical disease (NTD) associated with severe health consequences, including death. Hibiscus tiliaceus L., from the Malvaceae family, is used traditionally in Kisangani, Democratic Republic of the Congo (DRC), to alleviate symptoms of SCD. However, the specific phytochemicals responsible for the observed therapeutic effects remain unclear. This study aims to characterize the aqueous leaf extract of H. tiliaceus and assess its biological activity against sickle cell disease, including its antisickling, antioxidant, and anti-inflammatory effects. Using techniques such as TLC, HPLC-UV/DAD, LC-MS, and NMR, we identified kaempferol 3-O-rutinoside and rutin in the aqueous extract of H. tiliaceus leaves. Rutin exhibited potent antioxidant and anti-inflammatory activities, with IC50 values of 5 µg/mL and 2.5 µg/mL, respectively. Conversely, kaempferol 3-O-rutinoside demonstrated superior antisickling activity, normalizing sickled red blood cells with an IC50 < 12.5 µg/mL. Due to the pathophysiology of SCD, which involves the polymerization of red blood cells, which induces oxidative stress and an inflammatory response, this study suggests the importance of H. tiliaceus for the management of SCD. Additionally, the combined effect of molecules in H. tiliaceus will help in normalizing erythrocytes, inhibiting free radicals generated by early hemolysis, thus contributing to inflammatory processes reduction. This finding provides evidence and validates the traditional use of H. tiliaceus aqueous extract for the management of SCD.

The study of various microorganisms isolated from an Indian Ocean sponge, Scopalina hapalia ML-263, led to the selection of a promising Actinobacteria strain, Micromonospora sp. SH-82. Genomic analysis identified this strain as a new species, revealing the presence of 23 biosynthetic gene clusters (BGCs), some of which are associated with the synthesis of specialized metabolites such as polyketides deriving from polyketide synthases (PKSs). The strain was cultivated under favorable conditions for the production of bioactive molecules, resulting in the isolation and identification of seven microbial metabolites. Three of them are potentially novel, two erythronolides and one erythromycin, all characterized by a rare C10–C11 double bond. Some of these compounds also display atypical conformations, forming hemiacetals or spiroacetals. Their identification was achieved through detailed chemical analyses (NMR and ESI+-HRMS). A molecular networking approach was employed to assess the presence of potentially novel molecules in the microbial crude extract, supported by the identification of isolated molecules. Four molecules (1, 2, 3 and 5) were evaluated for their cytotoxic activities against cancer cell lines (HCT-116 and MDA-MB-231) and the immortalized retinal pigment epithelial RPE1 cells. No activity was observed in the latter, suggesting a lack of toxicity toward healthy cells. Moreover, megalomicin C1 (3), one of the isolated compounds, showed interesting antiplasmodial activity against Plasmodium falciparum 3D7, with an IC50 of 6.37 ± 2.99 µM.

Shea butter (SB) is a raw material fat obtained from Vitellaria paradoxa C.F. Gaertn kernels. We investigated the direct and indirect protective effects of 10 traditional and industrial SBs and their polar extracts on cell-free systems using ABTS and DPPH radical scavenging assays as well as on singlet oxygen (1O2) produced by Rose Bengal (RB) photosensitization. Their effects against RB-induced HaCaT cell phototoxicity were also explored. A spectrophotometric assay and HPLC were performed to quantify and identify phenolic content, which was between 14.16 and 82.99 ppm pyrogallol equivalent. These variations could be due to the SB origin and extraction process. These polar fractions exhibited moderate DPPH and strong ABTS radical-scavenging activity. By applying the UV–visible technique, we demonstrated that SBs and their phenolic compounds behave as 1O2 quenchers in a dose-dependent manner. Moreover, using a UVR-like model after the irradiation of RB, both polar extracts and crude SB exhibited photoprotective effects, highlighting the indirect protective action. In acellular and cellular models, SB and its polar extracts can act as a free radical scavenger against reactive oxygen species and 1O2 quenchers. Due to the maximum absorbance of SB at 280 nm and the antioxidant effect of 1O2 quenching, SB polar extracts exhibit photoprotective properties.

With over 3000 representatives, the monoterpene indole alkaloids (MIAs) class is among the most diverse families of plant natural products. The MS/MS spectral space exploration of these complex compounds using chemoinformatic and computational mass spectrometry tools offers a valuable opportunity to extract and share chemical insights from this emblematic family of natural products (NPs). In this work, we first present a substantially updated version of the MIADB, a database now containing 422 MS/MS spectra of MIAs that has been uploaded to the GNPS library versus 172 initial entries. We then introduce an innovative workflow that leverages hundreds of fragmentation spectra to support the FAIRification, extraction and dissemination of chemical knowledge. This workflow aims at the extraction of spectral patterns matching finely defined MIA skeletons. These extracted signatures can then be queried against complex biological extract datasets using MassQL. By applying this strategy to an LC-MS/MS dataset of 75 plant extracts, our results demonstrated the efficiency of this approach in identifying the diversity of MIA skeletons present in the analyzed samples. Additionally, our work enabled the digitization of structural data for diverse MIA skeletons by converting them into machine-readable formats and thereby enhancing their dissemination for the scientific community. Scientific contribution A comprehensive investigation of the monoterpene indole alkaloid chemical space, aiming to highlight skeleton-dependent fragmentation similarity trends and to generate valuable spectrometric signatures that could be used as queries.

Although primula species are abundant in phytochemicals, little is known about their pharmacological characteristics, especially those of endemic species. The purpose of this study was to examine the biological activity and phytochemical composition of the Georgian endemic Primula saguramica,as well as Primula macrocalyx, and the Caucasian endemic Primula woronowii. Folin–Ciocalteu and aluminum chloride colorimetric assays were used to measure the total phenolic content (TPC) and total flavonoid content (TFC), respectively. Cytotoxicity was assessed using the MTT assay in RAW 264.7 murine macrophages (ATCC-TIB-71), A549 lung adenocarcinoma cell line (ATCC-CCL-185), U87MG human glioblastoma cell line (ATCC-HTB-14), and NIH/3T3 - murine fibrobalst cell line (ATCC-CRL-1658), in addition to the hemolysis assay on human erythrocytes. The ABTS radical scavenging assay was used to confirm the antioxidant activity. An individual compound from the most active fraction (P.w.3, 50% methanolic fraction of P. woronowii) was isolated and identified using preparative high-performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR). For the quantitative determination of the individual compound in crude extracts and fractions, HPLC analysis was carried out, and its predicted pharmacological profile was evaluated using an in silico approach. P.w.3 had the highest TPC and TFC content (41.84 ± 0.08 mg GAE/g and 36.02 ± 0.2 mg QE/g) among all studied samples. It did not exhibit cytotoxic or hemolytic activity; however, it revealed potent antioxidant capacity (IC₅₀ = 7.35 ± 0.23 µg/mL), which was significantly stronger than clitorin (13.25 ± 0.24 µg/mL) and the crude extract P.w.1 (42.01 ± 0.21 µg/mL). A flavonoid glycoside, clitorin (kaempferol 3-O-(2'',6''-di-O-α-L-rhamnopyranosyl)-β-D-glucopyranoside), was identified from P.w.3 Clitorin content in P.w.3 was approximately ~5-fold higher (16.93 ± 1.16%, RSD) compared with P.w.1 (2.86 ± 2.5%, RSD). An in silico analysis showed that clitorin may have multifunctional biological activities, such as antioxidant, anti-inflammatory, antiproliferative, and vasoprotective effects. P.w.3 contained abundant bioactive flavonoids and exhibited strong antioxidant properties, without displaying cytotoxic or hemolytic effects. The isolated compound, clitorin, may serve as a lead in the discovery of phytotherapeutics for oxidative stress associated diseases.