Rapid Determination of Phenolics,Flavonoids, and Antioxidant Properties of Physalis ixocarpa Brot. ex Hornem. and Physalis angulata L. by Infrared Spectroscopy and Partial Least Squares

Physalis ixocarpa Brot. ex Hornem. and Physalis angulata L. are two edible species of the family Solanaceae, which have an important variety of antioxidant compounds present in their roots, stems, leaves, calyces, and fruits. This work reports the development of multivariate models based on the use of partial least square (PLS) analysis and Fourier transform infrared (FTIR) spectroscopy for the quantitative determination of total phenolics, total flavonoids, free radical scavenging activity, total antioxidant capacity, and reducing power in the extracts of roots, stems, and leaves of both P. ixocarpa and P. angulata. Standard chromatographic and colorimetric techniques were used to determine the quantitative actual values (references) in the extracts, which served as input data to develop the multivariate PLS models. Optimized FTIR-PLS models were realized by cross-validation procedures, obtaining the determination coefficients for prediction between 0.792 and 0.905 for P. ixocarpa, and between 0.756 and 0.893 for P. angulata. In this form, FTIR spectroscopy with multivariate analysis could represent a versatile tool to evaluate quantitatively concentrations of bioactive compounds and antioxidant properties in the extracts of both species, requiring a very short time at low cost.     

Eco-friendly synthesis of gold nanoparticles using fruit extracts and in vitro anticancer studies

Gold nanoparticles are biocompatible and are having several applications in biomedical Sciences and Engineering. Integration of nanoscience in medicine leads to the development of biomedical products that helps the Society in a faster and safer manner. In the present research work, bioreduction and biofunctionalization of gold nanoparticles are performed with fruit extracts of Aegle marmelos, Eugenia jambolana and soursop. The nanoparticles are characterized using UV–Vis spectroscopy, Transmission Electron Microscopy, Fourier Transform Infrared Spectroscopy and Zeta potentiometer. The qualitative phytochemical analysis of the fruit extracts shows the presence of alkaloids, amino acid, flavonoids, phenol, proteins, tannin, reducing sugars and total Sugars. The in vitro anticancer activity was confirmed by MTT assay on the human breast cancer cell line MCF-7 at different concentrations. The flavonoids present in the fruit extracts are potential reducing agent which is responsible for the formation of gold nanoparticles. Stabilization of gold nanoparticles are performed by the carboxylate group present in the proteins. Also, the nanoparticles are held apart from each other by the electrostatic repulsions that exist due to the presence of like charges surrounding the gold nanoparticles. This study proves that the fruit extracts can be used for the synthesis and stabilization of gold nanoparticles. Further, the engineered nanoparticles capped with bioactive compounds are potential anticancer agents against breast cancer cell line MCF-7.     

Cytotoxic withanolides from Physalis angulata

A new withanolide (1), physagulide P, together with five known withanolides (2–6), was isolated from the aerial parts of Physalis angulata L. The structure of new compound was elucidated on the basis of extensive spectroscopic techniques, including 1D, 2D NMR and HRESIMS. The activity screening indicated that compound 1 showed significant cytotoxicities against the human osteosarcoma cell line MG-63, HepG-2 hepatoma cells and breast cancer cells MDA-MB-231 with the IC₅₀ value of 3.50, 4.22 and 15.74 μM.     

Chemical Composition and Biological Activity of Argentinian Propolis of Four Species of Stingless Bees

The chemical composition of propolis of four species of stingless bees (SLBs) from Argentina was determined, and its antibacterial and anticancer activity was evaluated on selected types of microbes and cancer cell lines. Volatile secretions of all propolis samples are formed by 174 C₂–C₁₅ organic compounds, mainly mono- and sesquiterpenes and their derivatives. The chromatograms of ether extracts showed 287 peaks, of which 210 were identified. The most representative groups in the extracts of various propolis samples were diterpenoids (mainly resin acids), triterpenoids and phenolic compounds: long-chain alkenyl phenols, resorcinols and salicylates. The composition of both volatile and extractive compounds turned out to be species-specific; however, in both cases, the pairwise similarity of the propolis of Scaptotrigona postica and Tetragonisca fiebrigi versus that of Tetragona clavipes and Melipona quadrifasciata quadrifasciata was observed, which indicated the similarity of the preferences of the respective species when choosing plant sources of resin. The composition of the studied extracts completely lacked flavonoids and phenolcarboxylic acids, which are usually associated with the biological activity and medicinal properties of propolis. However, tests on selected microbial species and cancer cell lines showed such activity. All propolis samples tested against Paenibacillus larvae, two species of Bacillus and E. coli showed biofilm inhibition unrelated to the inhibition of bacterial growth, leading to a decrease in their pathogenicity. Testing the anticancer activity of ether extracts using five types of cell cultures showed that all four types of propolis studied inhibit the growth of cancer cells in a dose- and time-dependent manner. Propolis harvested by T. clavipes demonstrated the highest cytotoxicity on all tested cell lines.     

Rhamnus pallasii subsp. sintenisii fruit,leaf, bark and root: Phytochemical profiles and biological activities

The genus Rhamnus has received a lot of interest as a source of phenolic chemicals. There have been no reports on the phytochemicals and biological activities of R. pallasii subsp. sintenisii various morphological components (fruit, leaf, bark, and root) in Iran to yet. Two crude ether petroleum (EP) and hydro-methanolic (HM) extracts were obtained from the separate parts. The antioxidant and antibacterial capabilities of the extracts, as well as their phytochemical screening (total phenolic, flavonoid, phenolic acid, and anthocyanin concentrations), were measured. Furthermore, the phytochemical profiles of EP and HM extracts were determined using GC–MS and LC-ESI–MS, respectively. LC-ESI-MS detected 59 chemicals in HM extracts, including flavonoids (62.71 %), phenolic acids (10.16 %), and anthraquinones (16.94 %). Furthermore, the predominant group components in EP extracts examined by GC–MS were fatty acids (58.82%), phenolic compounds (49.28%), and hydrocarbons (35.15 to 59.45 %). In terms of biological testing (DPPH radical scavenging and anti-bacterial activity), all examined extracts, particularly the fruit, had the highest activities in both assays (IC₅₀: 7.52 to 22.39 µg/ml and MIC: 0.39 to 3.12 mg/ml), owing to their high phenolic content. As a result, individual morphological elements of the species might be thought of as natural antioxidant and antibacterial agents.     

Analysis of Plant–Plant Interactions Reveals the Presence of Potent Antileukemic Compounds

A method to identify anticancer compounds in plants was proposed based on the hypothesis that these compounds are primarily present in plants to provide them with an ecological advantage over neighboring plants and other competitors. According to this view, identifying plants that contain compounds that inhibit or interfere with the development of other plant species may facilitate the discovery of novel anticancer agents. The method was developed and tested using Magnolia grandiflora, Gynoxys verrucosa, Picradeniopsis oppositifolia, and Hedyosmum racemosum, which are plant species known to possess compounds with cytotoxic activities. Plant extracts were screened for growth inhibitory activity, and then a thin-layer chromatography bioautography assay was conducted. This located the major antileukemic compounds 1, 2, 4, and 5 in the extracts. Once the active compounds were located, they were extracted and purified, and their structures were determined. The growth inhibitory activity of the purified compounds showed a significant correlation with their antileukemic activity. The proposed approach is rapid, inexpensive, and can easily be implemented in areas of the world with high biodiversity but with less access to advanced facilities and biological assays.     

Determination of Cytotoxic Activity of Sanguinaria canadensis Extracts against Human Melanoma Cells and Comparison of Their Cytotoxicity with Cytotoxicity of Some Anticancer Drugs

Melanoma is an enormous global health burden, and should be effectively addressed with better therapeutic strategies. Therefore, new therapeutic agents are needed for the management of this disease. The aim of this study was the investigation of cytotoxic activity of some isoquinoline alkaloid standards and extracts obtained from Sanguinaria canadensis—collected before, during, and after flowering—against three different human melanoma cells (A375, G361, SK-MEL-3). The cytotoxicity of these extracts was not previously tested on these melanoma cell lines. Determination of alkaloid contents was performed by HPLC-DAD using Polar RP column and mobile phase containing acetonitrile, water, and 1-butyl-3-methylimidazolium tetrafluoroborate. The cytotoxicity of alkaloid standards was investigated by determination of cell viability and calculation of IC₅₀ values. Significant differences were observed in the alkaloids content and cytotoxic activity of the extracts, depending on the season of collection of the plant material. In the Sanguinaria canadensis extracts high contents of sanguinarine (from 4.8543 to 9.5899 mg/g of dry plant material) and chelerythrine (from 42.7224 to 6.8722 mg/g of dry plant material) were found. For both of these alkaloids, very high cytotoxic activity against the tested cell lines were observed. The IC₅₀ values were in the range of 0.11–0.54 µg/mL for sanguinarine and 0.14 to 0.46 µg/mL for chelerythrine. IC₅₀ values obtained for Sanguinaria canadensis extracts against all tested cell lines were also very low (from 0.88 to 10.96 µg/mL). Cytotoxic activity of alkaloid standards and Sanguinaria canadensis extracts were compared with the cytotoxicity of anticancer drugs—etoposide, cisplatin, and hydroxyurea. In all cases except the one obtained for cisplatin against A375, which was similar to that obtained for Sanguinaria canadensis after flowering against the same cell line, IC₅₀ values obtained for anticancer drugs were higher than the IC₅₀ values obtained for sanguinarine, chelerythrine, and Sanguinaria canadensis extracts. Our results showed that Sanguinaria canadensis extracts and isoquinoline alkaloids, especially sanguinarine and chelerythrine, could be recommended for further in vivo experiments in order to confirm the possibility of their application in the treatment of human melanomas.     

Antibacterial and Antioxidant Activity of Keladi Tikus Leaves Extract (Typhonium Flagelliforme) (Lodd) Blume

Keladi tikus (Typhonium flagelliforme (Lodd) Blume is a plant that has many benefits in - health such as anticancer, anti-inflammatory, analgesic and antihepatotoxic. This study aims to determine the antibacterial and antioxidant activity of different extracts of leaves T. flagelliforme. The agar diffusion method used in the antibacterial activity; DPPH, the FTC and the TBA method used in the antioxidant activity. The results showed the ethyl acetate, n-butanol and water fraction had antibacterial activity against Bacillus subtilis and Pseudomonas aeruginosa, while the n-hexane fraction had no activity against bacteria tested. The results of antioxidant activity by DPPH method, the FTC and the TBA showed ethyl acetate fraction was the most active (IC₅₀ = 56.32 ppm) fraction among others.     

Discovery of Leptulipin,a New Anticancer Protein from theIranian Scorpion,Hemiscorpius lepturus

Cancer is one of the leading causes of mortality in the world. Unfortunately, the present anticancer chemotherapeutics display high cytotoxicity. Accordingly, the discovery of new anticancer agents with lower side effects is highly necessitated. This study aimed to discover an anticancer compound from Hemiscorpius lepturus scorpion venom. Bioactivity-guided chromatography was performed to isolate an active compound against colon and breast cancer cell lines. 2D electrophoresis and MALDI-TOF were performed to identify the molecule. A partial protein sequence was obtained by mass spectrometry, while the full-length was deciphered using a cDNA library of the venom gland by bioinformatics analyses and was designated as leptulipin. The gene was cloned in pET-26b, expressed, and purified. The anticancer effect and mechanism action of leptulipin were evaluated by MTT, apoptosis, and cell cycle assays, as well as by gene expression analysis of apoptosis-related genes. The treated cells displayed inhibition of cell proliferation, altered morphology, DNA fragmentation, and cell cycle arrest. Furthermore, the treated cells showed a decrease in BCL-2 expression and an increase in Bax and Caspase 9 genes. In this study, we discovered a new anticancer protein from H. lepturus scorpion venom. Leptulipin showed significant anticancer activity against breast and colon cancer cell lines.     

Phytochemical Analysis and Anti-Cancer Properties of Extracts of Centaurea castriferrei Borbás & Waisb Genus of Centaurea L

The Centaurea L. (Asteraceae) genus includes many plant species with therapeutic properties. Centaurea castriferrei Borbás & Waisb is one of the least known and least described plants of this genus. The aim of the study was the phytochemical analysis of water and methanol–water extracts (7:3 v/v) obtained from the aerial parts of the plant as well as evaluation of their anticancer activity. Quantitative determinations of phenolic compounds and flavonoids were performed, and the antioxidant potential was measured using the CUPRAC method. The RP-HPLC/DAD analysis and HPLC-ESI-QTOF-MS mass spectroscopy were performed, to determine the extracts’ composition. The antiproliferative activity of the obtained extracts was tested in thirteen cancer cell lines and normal skin fibroblasts using MTT test. Regardless of the extraction method and the extractant used, similar cytotoxicity of the extracts on most cancer cell lines was observed. However, the methanol–water extracts (7:3 v/v) contained significantly more phenolic compounds and flavonoids as well as showing stronger antioxidant properties in comparison to water extracts. Centaurea castriferrei Borbás & Waisb is a rich source of apigenin and its derivatives. In all tested extracts, chlorogenic acid and centaurein were also identified. In vitro research revealed that this plant may be a potential source of compounds with anticancer activity.     

Synthesis,antibacterial, anticancer and molecular docking studies of macrocyclic metal complexes of dihydrazide and diketone

The complexes of Co(II), Ni(II), Cu(II) and Zn(II) metal ions has been synthesized through template method by the condensation of succinic acid dihydrazide with 5-chloroisatin in alcoholic medium. Complexes were characterized by C H N analysis, molar conductance, thermal analysis, magnetic susceptibility, mass spectrometry, FTIR, EPR, ¹H NMR, UV–Visible spectroscopy. These studies suggest an octahedral geometry for all the complexes. The compounds were found active against B. subtilis and S. aureus and P. aeruginosa and E. coli bacteria. The Zn(II) complex showed significant anticancer activity against Squamous Cell Carcinoma cells tested by the MTT assay method. Molecular docking studies with EGFR tyrosine kinase were also carried out. All these results show that some of the synthesized compounds have remarkable antibacterial and anticancer property.     

Changes in Physicochemical,Free Radical Activity,Total Phenolic and Sensory Properties of Orange (Citrus sinensis L.) Juice Fortified with Different Oleaster (Elaeagnus angustifolia L.) Extracts

In Iran and other parts of Western Asia, the oleaster (Elaeagnus angustifolia L.) fruit is processed in the dried powdery form, and in recent times, increasingly applied/sprinkled in fruit juices such as those made from oranges (Citrus sinensis L.). To our best knowledge, the effectiveness of oleaster fruit extract in fortifying the orange juice has not yet been reported and the knowledge of this will greatly benefit the consumers, particularly those around the Western Asia region. This current work, therefore, investigated the changes in physicochemical, free radical activity, total phenolic compounds, and sensory properties of orange juice fortified with different oleaster fruit extracts. The orange juice mix formulation comprised different concentrations (5, 10, 15, 20, and 25%) of oleaster (alcoholic, aqueous, and hydro-alcoholic) extracts. The control comprised orange concentrate (4% w/v), sugar (8.5% w/v), and citric acid (0.1% w/v) brought to the desirable volume with water. As the free radical activity depicted the antioxidant properties, the physicochemical aspects of this work involved the determinations of Brix, density, ash, pH, total acidity, sucrose, and total sugar, whereas the sensory aspects involved the determinations of color and taste. Whilst the aqueous oleaster 20 and 25% extracts produced notable physicochemical differences in the orange juice mix, both free radical activity, and phenolic compounds significantly increased (p < 0.05) after 30 days despite resembling (p > 0.05) those of control at day 1. More so, the increases in aqueous, alcoholic, and hydro-alcoholic oleaster extracts would decrease (p < 0.05) the sensory color and taste of the orange juice mix in this study.     

Mahonia aquifolium Extracts Promote Doxorubicin Effects against Lung Adenocarcinoma Cells In Vitro

Mahonia aquifolium and its secondary metabolites have been shown to have anticancer potential. We performed MTT, scratch, and colony formation assays; analyzed cell cycle phase distribution and doxorubicin uptake and retention with flow cytometry; and detected alterations in the expression of genes involved in the formation of cell–cell interactions and migration using quantitative real-time PCR following treatment of lung adenocarcinoma cells with doxorubicin, M. aquifolium extracts, or their combination. MTT assay results suggested strong synergistic effects of the combined treatments, and their application led to an increase in cell numbers in the subG1 phase of the cell cycle. Both extracts were shown to prolong doxorubicin retention time in cancer cells, while the application of doxorubicin/extract combination led to a decrease in MMP9 expression. Furthermore, cells treated with doxorubicin/extract combinations were shown to have lower migratory and colony formation potentials than untreated cells or cells treated with doxorubicin alone. The obtained results suggest that nontoxic M. aquifolium extracts can enhance the activity of doxorubicin, thus potentially allowing the application of lower doxorubicin doses in vivo, which may decrease its toxic effects in normal tissues.     

Phytochemical Characterization and Bioactivity of Asparagus acutifolius: A Focus on Antioxidant,Cytotoxic, Lipase Inhibitory and Antimicrobial Activities

The phytochemical composition of leaves, stems, pericarps and rhizomes ethanolic extracts of Asparagus acutifolius were characterized by HPLC-DAD-MS. A. acutifolius samples contain at least eleven simple phenolics, one flavonon, two flavonols and six steroidal saponins. The stem extracts showed the highest total phenolic acid and flavonoid contents, where cafeic acid and rutin were the main compounds. No flavonoids were detected in the leaf, pericarp or rhizome while caffeic acid and ferulic acid were the predominant. Steroidal saponins were detected in the different plant parts of A. acutifolius, and the highest contents were found in the rhizome extracts. The stem extracts exhibited the highest antioxidant activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) and the highest 2,2-azino-bis (3 ethylbenzothiazoline-6-sulphonic acid) (ABTS) scavenging activity was found in the pericarp extracts. The rhizome and leaf extracts showed a potent cytotoxic activity against HCT-116 and HepG2 cell lines. Moreover, the pericarp and rhizome extracts revealed a moderate lipase inhibitory activity. The leaf and rhizome extracts were screened for their antimicrobial activity against human pathogenic isolates. The leaf extract exhibited a powerful inhibitory activity against all the bacteria and fungi tested.     

Peptaibol-Containing Extracts of Trichoderma atroviride and the Fight against Resistant Microorganisms and Cancer Cells

Fighting resistance to antibiotics and chemotherapeutics has brought bioactive peptides to the fore. Peptaibols are short α-aminoisobutyric acid-containing peptides produced by Trichoderma species. Here, we studied the production of peptaibols by Trichoderma atroviride O1 and evaluated their antibacterial and anticancer activity against drug-sensitive and multidrug-resistant bacterium and cancer cell lines. This was substantiated by an analysis of the activity of the peptaibol synthetase-encoding gene. Atroviridins, 20-residue peptaibols were detected using MALDI-TOF mass spectrometry. Gram-positive bacteria were susceptible to peptaibol-containing extracts of T. atroviride O1. A synergic effect of extract constituents was possible, and the biolo-gical activity of extracts was pronounced in/after the peak of peptaibol synthetase activity. The growth of methicillin-resistant Staphylococcus aureus was reduced to just under 10% compared to the control. The effect of peptaibol-containing extracts was strongly modulated by the lipoteichoic acid and only slightly by the horse blood serum present in the cultivation medium. Peptaibol-containing extracts affected the proliferation of human breast cancer and human ovarian cancer cell lines in a 2D model, including the multidrug-resistant sublines. The peptaibols influenced the size and compactness of the cell lines in a 3D model. Our findings indicate the molecular basis of peptaibol production in T. atroviride O1 and the potential of its peptaibol-containing extracts as antimicrobial/anticancer agents.     

Rosa spp. Extracts as a Factor That Limits the Growth of Staphylococcus spp. Bacteria,a Food Contaminant

Due to their richness of bioactive substances, rose hips are a valuable raw material for obtaining extracts with potential antimicrobial activity. The aim of the study was to determine the antagonistic potential of whole pseudo-fruit and flesh extracts of three Rosa sp. varieties against Staphylococcus spp. bacteria isolated as food contaminants. The biological material in this study consisted of seven strains of bacteria from the genus Staphylococcus. Two strains—Staphylococcus aureus ATCC 25923 and Staphylococcus epidermidis DSMZ 3270—were used as reference strains. The other five strains were food-derived isolates—S. epidermidis A5, S. xylosus M5, S. haemolyticus M6, S. capitis KR6, and S. warneri KR2A. The material was the pseudo-fruits of Rosa canina, Rosa pomifera Karpatia, and Rosa rugosa. The polyphenols were extracted from the fleshy part and the whole pseudo-fruit for all rose varieties. The tested preparations differed significantly in their polyphenol composition. The sum of polyphenols ranged from 28 862 to 35 358 mg/100 g of lyophilisate. The main groups of polyphenols found in the preparations were flavanols and ellagitannins. All of the tested extracts inhibited the growth of staphylococci at a concentration of 500 mg/mL. Rosa rugosa fruit extract showed the strongest antimicrobial properties among the studied extracts. For all the strains, the growth inhibition had a diameter of 20.3–29.0 mm. Moreover, six out of the seven tested strains showed the highest inhibition with the use of this extract. The MIC of rose extracts was in the range of 3.125–500 mg/mL and was strictly dependent on the bacterial species, the species of the rose, and the part of the fruit from which the extract was obtained. Correlations were assessed between the main groups of polyphenols in the extracts and their inhibition of bacterial growth. In the case of pseudo-fruit extracts, the inhibitory effect on bacterial growth positively correlated with the content of ellagitannins, and this effect was observed for almost all the tested strains. The results presented herein follow the current trend of minimising the use of chemical preservatives in food; from this point of view, rose extracts are very promising.     

Phenolic Profile,EPR Determination,and Antiproliferative Activity against Human Cancer Cell Lines of Anthyllis vulneraria Extracts

In the current work, the leaf and flower extracts of Anthyllis vulneraria were evaluated for their chemical characterization using HPLC-MS and for their radical scavenging capacity toward methoxy radicals produced by a Fenton-type reaction using an electron paramagnetic resonance (EPR) spectroscopy assay. The in vitro antiproliferative activity of these extracts against several human-derived cancer cells (breast: MCF-7; cervical: HeLa; hepatocellular: HepG2) was also evaluated. The results showed that the Anthyllis vulneraria leaf extract was characterized by 17 different phenolic compounds, among which phenolic acids were the most abundant, while its flower extract exhibited higher contents of flavonoids. Furthermore, Anthyllis vulneraria extracts demonstrated a potent radical scavenging activity against methoxy radicals. Both extracts also significantly reduced the viability of the different cancer cell lines. The results of the current study suggested that Anthyllis vulneraria extracts are a promising source of antioxidant compounds with health benefits and pointed to their potential use for treating cancer and developing novel therapeutic agents.     

Antitumor Potential of Annona muricata Linn. An Edible and Medicinal Plant in Mexico: In Vitro,In Vivo,and Toxicological Studies

Annona muricata (Am) is a plant used in traditional Mexican medicine to treat cancer. In this study, ethanol extracts of Am collected in Acapulco and Tecpan from Guerrero state were evaluated orally on Balb/c mice inoculated with 4T1 cells, for cytotoxic activity (CA) on 4T1 cells, in brine shrimp lethality assay (BSLA), and for acute oral toxicity in mice. In addition, ethanol extracts were subjected to high-performance liquid chromatography (HPLC) with diode array detection. Results showed that the extracts collected in December in Acapulco (AcDe) and Tecpan (TeDe) exhibited the most significant antitumor and cytotoxic activity. In the BSLA, the most important effect was observed in the extracts from Acapulco and Tecpan collected in June (AcJu) and August (TeAg), respectively. The samples from Acapulco (AcJu, and AcAg) and Tecpan (TeJu and TeAg) showed the highest toxicity. The analysis of the extracts, AcDe and TeDe, by HPLC revealed that flavonoids, rutin, narcissin, and nicotinflorin were the major components. These findings suggest that extracts from Am collected in Acapulco and Tecpan in the month of December may be an important source to obtain flavonoid glycosides with anticancer potential specifically against breast cancer. This also supports the use of Am to treat cancer in Mexican traditional medicine.     

Potential Anticancer Activity of the Furanocoumarin Derivative Xanthotoxin Isolated from Ammi majus L. Fruits: In Vitro and In Silico Studies

Ammi majus L., an indigenous plant in Egypt, is widely used in traditional medicine due to its various pharmacological properties. We aimed to evaluate the anticancer properties of Ammi majus fruit methanol extract (AME) against liver cancer and to elucidate the active compound(s) and their mechanisms of action. Three fractions from AME (Hexane, CH₂Cl₂, and EtOAc) were tested for their anticancer activities against HepG2 cell line in vitro (cytotoxicity assay, cell cycle analysis, annexin V-FITC apoptosis assay, and autophagy efflux assay) and in silico (molecular docking). Among the AME fractions, CH₂Cl₂ fraction revealed the most potent cytotoxic activity. The structures of compounds isolated from the CH₂Cl₂ fraction were elucidated using ¹H- and ¹³C-NMR and found that Compound 1 (xanthotoxin) has the strongest cytotoxic activity against HepG2 cells (IC₅₀ 6.9 ± 1.07 µg/mL). Treating HepG2 cells with 6.9 µg/mL of xanthotoxin induced significant changes in the DNA-cell cycle (increases in apoptotic pre-G1 and G2/M phases and a decrease in the S-phase). Xanthotoxin induced significant increase in Annexin-V-positive HepG2 cells both at the early and late stages of apoptosis, as well as a significant decrease in autophagic flux in cancer compared with control cells. In silico analysis of xanthotoxin against the DNA-relaxing enzyme topoisomease II (PDB code: 3QX3) revealed strong interaction with the key amino acid Asp479 in a similar fashion to that of the co-crystallized inhibitor (etoposide), implying that xanthotoxin has a potential of a broad-spectrum anticancer activity. Our results indicate that xanthotoxin exhibits anticancer effects with good biocompatibility toward normal human cells. Further studies are needed to optimize its antitumor efficacy, toxicity, solubility, and pharmacokinetics.     

Phytochemical Profiles,Antioxidant, Cytotoxic,and Anti-Inflammatory Activities of Traditional Medicinal Plants: Centaurea pichleri subsp. pichleri,Conyza canadensis,and Jasminum fruticans

Centaurea pichleri subsp. pichleri, Conyza canadensis, and Jasminum fruticans are traditionally used plants grown in Turkey. Methanol extracts were obtained from these plants and pharmacological activity studies and phytochemical analyses were carried out. To evaluate the phytochemical composition, spectrophotometric and chromatographic techniques were used. The extracts were evaluated for antioxidant activity by DPPH^●, ABTS^●+ radical scavenging, and FRAP assays. The cytotoxic effects of the extracts were investigated on DU145 prostate cancer and A549 lung cancer cell lines. The anti-inflammatory effects of extracts were investigated on the NO amount, TNF-α, IFN-γ, and PGE 2 levels in lipopolysaccharide-stimulated Raw 264.7 cells. The richest extract in terms of phenolic compounds (98.19 ± 1.64 mg_GAE/g_extract) and total flavonoids (21.85 ± 0.64 mg_CA/g_extract) was identified as C. pichleri subsp. pichleri methanol extract. According to antioxidant activity determinations, the C. pichleri subsp. pichleri extract was found to be the most active extract. Finally, the C. pichleri subsp. pichleri methanol extract was revealed to be the most effective inhibitor of viability in the cytotoxic activity investigation, and the extract with the best anti-inflammatory action. The findings point to C. pichleri subsp. pichleri as a promising source of bioactive compounds in the transition from natural sources to industrial uses, such as new medications, cosmeceuticals, and nutraceuticals.