A review of the ethnomedicinal, phytochemical, and pharmacological properties of the Ferulago genus based on Structure-Activity Relationship (SAR) of coumarins.

BACKGROUND: The Ferluago W.D.J. Koch genus includes 48 accepted perennial herbs that are distributed in the Mediterranean region, Southeast Europe, Central and Middle East of Asia. These plants are widely used in folk and conventional medicine due to their biological benefits such as anti-microbial, anti-inflammatory, anti-cancer, and immunomodulatory properties. Conducting a comprehensive review based on the structure activity relationships (SARs) of the coumarins, which has not been previously documented, can lead to a better insight into the genus Ferulago and its beneficial therapeutic activities. METHODS: This review covers literature from 1969 to 2023, were collected from various scientific electronic databases to review phytochemical, pharmacological, and ethnopharmacological data of Ferulago species, as well as latest information on the SAR of reported coumarins from this genus. RESULTS: Phytochemical studies showed that the biological actions of this genus are mediated by the reported specialized metabolites, such as coumarins and flavonoids. Simple coumarins, prenylated coumarins, furanocoumarins, and pyranocoumarins are the largest subclasses of coumarins found in diverse Ferulago species, which have discussed the biological effects of them with a focus on the Structure-Activity Relationship (SAR). For example, prenylated coumarins have shown potential leishmanicidal and anti-neuropsychiatric effects when substituted with a prenyl group at the 7-hydroxy, as well as the C6 and C8 positions in their scaffold. Similarly, furanocoumarins exhibit varied biological activities such as anti-inflammatory, anti-proliferative, and anti-convulsant effects. Modifying substitutions at the C5 and C6 positions in furanocoumarins can enhance these activities. CONCLUSION: This study conducted a comprehensive review of all available information on the phytochemical and pharmacological characteristics of Ferulago species. Given the high occurrence of coumarins in this genus, which exhibit potential anti-Alzheimer and anti-microbial properties, it presents promising new therapeutic avenues for addressing these common issues. Further investigation is needed to understand the molecular-level mechanisms of action and to explore their clinical applications.

α-Glucosidase inhibition assay of galbanic acid and it amide derivatives: New excellent semi-synthetic α-glucosidase inhibitors.

α-Glucosidase inhibitory activity of galbanic acid and its new amide derivatives 3a-n were investigated. Galbanic acid and compounds 3a-n showed excellent anti-α-glucosidase activity with IC50 values ranging from 0.3 ± 0.3 µM to 416.0 ± 0.2 µM in comparison to positive control acarbose with IC50 value of = 750.0 ± 5.6. In the kinetic study, the most potent compound 3h demonstrated a competitive mode of inhibition with Ki = 0.57 µM. The interaction of the most potent compound 3h with the α-glucosidase was further elaborated by in vitro Circular dichroism assessment and in silico molecular docking and Molecular dynamics studies. Compound 3h was also non-cytotoxic on human normal cells. In silico study on pharmacokinetics and toxicity profile of the most potent galbanic acid derivatives demonstrated that these compounds are valuable lead compounds for further study in order to achieve new anti-diabetic agents.

New antiprotozoal sesquiterpene derivatives from Dorema glabrum Fisch & C.A.Mey.

Seven undescribed sesquiterpene derivatives, Azerins A-G (3-6, 8, 14 and 15), three known sesquiterpene phenols, kopetdaghin A (1), kopetdaghin B (2) and latisectin (7), together with five known sesquiterpene coumarins (9-13), were isolated from the roots of Dorema glabrum. The structures were elucidated by comprehensive 1D- and 2D-NMR spectral analysis as well as HR-ESI-MS. Compounds were assessed for their in vitro antiprotozoal activity against Trypanosoma brucei rhodesiense, T. cruzi, Leishmania donovani, and Plasmodium falciparum. Cytotoxic potentials of the compounds were also tested on L6 rat skeletal myoblasts. Azerin G (15) showed a potent preferential growth inhibitory activity against T. b. rhodesiense with IC50 value of 0.01 µM and selectivity index of 329. Compounds 1, 4, 7 and 8 were also found as the most active compounds with selective growth inhibitory effects toward P. falciparum with selectivity indices ranging from 11.6 to 16.7 (IC50: 1.8-24.6 µM).

Cytotoxic Activity and Phytochemical Analysis of Artemisia haussknechtii Boiss.

Cytotoxic activity of crude extract and fractions (petroleum ether, dichloromethane, and n-butanol) of Artemisia haussknechtii aerial parts was investigated by MTT assay. Dichloromethane fraction showed the highest cytotoxic effect on MCF-7 cell line (IC50 = 297.17 ± 7.99 µg/mL). Phytochemical analysis of the most effective fraction was carried out using normal phase column chromatography (CC) to get eight sub-fractions (A-H). Thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) were used for further purification. Four known compounds with cytotoxic effects on cancer cell lines were isolated from the most active fraction, including 5-Hydroxy-3',4',6,7-tetramethoxyflavone (eupatilin 7-methyl ether), 5-hydroxy 3,3',4',6,7-pentamethoxy-flavone (artemetin), 6-methoxy-7-hydroxycoumarin (scopoletin), and methyl caffeate. Structure elucidation of isolated compounds was done using spectroscopic techniques, including ESIMS and 1D-NMR (1H and 13C). Cytotoxic activity of A. haussknechtii is probably due to coumarin and flavonoid compounds.

Free Radical Scavenging Principles of Salvia reuterana Boiss. Aerial Parts.

Salvia reuterana Boiss. is an aromatic perennial plant traditionally used for its anxiolytic and sedative properties. In the present study, various fractions and essential oil of S. reuterana aerial parts were investigated to find its free radical scavenging principles. Hydroalcoholic fraction with IC50 value of 112.6 ±3.2 µg mL-1 in DPPH assay demonstrated the highest free radical scavenging activity and was selected to further phytochemical investigation. RP-18 and Sephadex LH-20 column chromatography of the hydroalcoholic fraction resulted in the isolation and structural elucidation of four phenolic derivatives, including apigenin-7-O-ß-D-glucopyranoside (1), luteolin-7-O-ß-D-glucopyranoside (2), rosmarinic acid (3), and luteolin (4). Isolated compounds showed potent free radical scavenging activities (5.1-34.2 µg mL-1), compared with BHT (21.30 ± 1.9 µg mL-1). Twenty four compounds were also identified in GC-MS analysis of the plant essential oil, of which benzyl benzoate (26.64%), n-hexyl benzoate (22.99%) and n-hexyl isovalerate (6.04%) were the main compounds. The results of the present study introduced S. reuterana as a valuable source of natural phenolic antioxidants which can be utilized in prevention of oxidative stress related diseases. Moreover, interesting composition of S. reuterana essential oil, dominated by non-terpenes compounds (76.17%) especially aromatic derivatives, make it an appropriate candidate for more detailed studies.

1, 5-dicaffeoylquinic acid, an α-glucosidase inhibitor from the root of Dorema ammoniacum D. Don.

BACKGROUND AND PURPOSE: Dorema ammoniacum D. Don (Apiaceae family) is a perennial plant whose oleo- gum resin is used as a natural remedy for various diseases, especially chronic bronchitis, and asthma. In the present study, hydromethanolic extract of D. ammoniacum root was subjected to phytochemical analyses and α-glucosidase inhibitory potentials of the isolated compounds were assessed. EXPERIMENTAL APPROACH: Silica gel (normal and reversed phases) and Sephadex® LH-20 column chromatographies were used for the isolation and purification of the compounds. Structures of the compounds were characterized by 1D and 2D nuclear magnetic resonance (NMR) techniques. All the isolated compounds were assessed for their in vitro α-glucosidase inhibitory activity in comparison with acarbose, a standard drug. FINDINGS/RESULTS: Two phloroacetophenone glycosides; echisoside (1) and pleoside (2), along with dihydroferulic acid-4-O-ß-D-glucopyranoside (3), and ß-resorcylic acid (4), and two caffeoylquinic acid derivatives; chlorogenic acid (5) and 1, 5-dicaffeoylquinic acid (cynarin, 6) were isolated. Among the isolated compounds, the α-glucosidase inhibitory effect of 1,5-dicaffeoylquinic acid was found as 76.9% of the acarbose activity at 750 µM (IC50 value of acarbose). CONCLUSION AND IMPLICATIONS: Considerable α-glucosidase inhibitory effect of 1,5-dicaffeoylquinic acid makes it an appropriate candidate for further studies in the development of new natural antidiabetic drugs.

Cell Cycle Arrest and Apoptosis Induction of Phloroacetophenone Glycosides and Caffeoylquinic Acid Derivatives in Gastric Adenocarcinoma (AGS) Cells.

INTRODUCTION: In the present study, we analyzed anti-proliferative and apoptosis induction activity of five phenolic compounds: echisoside, pleoside, chlorogenic acid, 4,5-Di-O-caffeoylquinic acid, and cynarin on AGS (adenocarcinoma gastric) cell line. METHOD: These phenolic compounds were isolated from methanol extract of Dorema glabrum root. An MTT assay was conducted to evaluate the inhibitory effect on cancer cells. EB/AO staining was done to assess the mode of cell death and morphological changes of the cells' nuclei. Cell cycle distribution of the cells was analyzed by flow cytometry, and for further confirmation of the pathway, mRNA levels of apoptosis cascade players were quantified by qRT-PCR. RESULT: We found that echisoside, pleoside, chlorogenic acid, 4,5-Di-O-caffeoylquinic acid, and cynarin inhibited the proliferation of AGS cancer cells in vitro. Our data revealed that these compounds triggered morphological changes characteristic of apoptotic cell death. These compounds up-regulated bax and caspase3 expression and down-regulated cyclin D1, bcl2, VEGFA, c-myc and survivin. Moreover, cell population increased at the G1 phase, and a number of cells at the G2/M phase of the cell cycle decreased after treatment. CONCLUSION: All these data suggest that phenolic compounds have a cytotoxic effect on gastric cancer cells and could trigger apoptosis. Besides cytotoxic activity, they could potentially arrest the cell cycle at the G1 phase.

Bioactive coumarins from the roots and fruits of Ferulago trifida Boiss., an endemic species to Iran.

Phytochemical analysis of the Ferulago trifida Boiss. from Apiaceae family led to the isolation and identification of suberosin (1), isoimperatorin (2), prantschimgin (3), oxypeucedanin (4), oxypeucedanin methanolate (5), suberenol (6), 6-hydroxymethylherniarin (7), oxypeucedanin hydrate (8), ulopterol (9), bergapten (10), xanthotoxin (11), imperatorin (12) and grandivittin (13) from chloroform extracts of the roots (1-9) and fruits (1, 2, 8, 10-13) of this species. Oxypeucedanin methanolate and suberenol demonstrated a potent antioxidant power with 268.2 ± 5.4 and 251.2 ± 6.2 mmol FSE/100 g, respectively, compared by BHT (267.2 ± 4.2 mmol FSE/100 g) in FRAP method. The potent antibacterial effects were found for oxypeucedanin methanolate on S. epidermidis (IZ; 26 mm, MIC; 250 µg mL-1) an oxypeucedanin hydrate on K. pneumoniae (IZ: 21 mm, MIC: 250 µg mL-1). Moreover, suberosin showed higher preferential toxicity against MDA-MB-23 cells (IC50: 0.21 mM, SI: 5.0), in comparison with tamoxifen (IC50: 0.012 mM, SI: 2.45) in MTT assay.

A Survey on Phytochemical Composition and Biological Activity of Zygophyllum fabago from Iran.

Purpose: Zygophyllum fabago L. (Z. fabago) is a widespread perennial herb which is used as a medicinal plant in traditional medicine of Iran, Turkey and China. The present study was a survey on phytochemical constituents and biological activities of this plant. Methods: Methanolic extract of the roots was fractionated over a C-18 pre-packed cartridge (Sep-pak) and chromatographic separation was performed on a reversed-phase preparative HPLC. Structural elucidation of the isolated compounds was carried out using UV, 1H-NMR and 13C-NMR spectral analyses. Furthermore, the chemical compositions of the essential oil of the aerial parts were identified by GC-MS analysis. Antiproliferative and antioxidant activities of all extracts from aerials were determined by MTT and DPPH assays, respectively. Results: Phytochemical investigation on the plant roots led to the isolation and identification of two the 60% methanol-water Sep-pak fraction, a prenylated flavone glycoside, 6-C-prenyl-7-O-[ ß -D-4'''-O-acetyl-glucopyranosyl-(1'''→2'')-ß-D-glucopyranosyl] apigenin, which was named as a Zygocaperoside and also, other flavonoid, was named as the Isorhamnetin -3-O glucoside. None of the extracts showed antiproliferative effect against cancerous cells. However, among the extracts, methanolic extract indicated antioxidant activity. Moreover, essential oils of flowers and leaves of plant have high amounts of sesquiterpene hydrocarbons and diterpenoides. Conclusion: The results of present study introduce Z. fabago roots as a new source of flavonoid glycosides and suggest it as an appropriate candidate for further pharmacological studies.

Isolation of Phenolic Derivatives and Essential Oil Analysis of Prangos ferulacea (L.) Lindl. Aerial Parts.

Prangos ferulacea (L.) Lindl. (Apiaceae) is a medicinal plant distributed in Mediterranean regions, Caucasia and southwest of Asia. In the present study phytochemical constituents of the extract obtained from the aerial parts of P. ferulacea were investigated using various chromatographic and spectroscopic methods. Essential oil of the plant aerial parts was also analyzed using GC-MS. Five phenolic derivatives, isoimperatorin (1), ferudenol (2), caffeic acid glucosyl ester (3), isorhamnetin-3-O-ß-D-glucopyranoside (4) and quercetin-3-O-ß-D-glucopyranoside (5) were isolated from the aerial parts of P. ferulacea and their structures were elucidated using 1H-NMR, 13C-NMR, EI-MS and UV spectral analyses. Twenty-seven compounds were also identified in the essential oil of plant aerial parts, of which ß-pinene (43.1%), α-pinene (22.1%) and -δ3-carene (16.9%) were characterized as main compounds. The results of this study introduce P. ferulacea as a source of potentially bioactive phenolic compounds and suggest it as an appropriate candidate for further studies.

Induction of Apoptosis and Cell Cycle Arrest by Dorema Glabrum Root Extracts in a Gastric Adenocarcinoma (AGS) Cell Line

Objective: Dorema glabrum Fisch. & C.A. Mey is a perennial plant that has several curative properties. Anti-proliferative activity of seeds of this plant has been demonstrated in a mouse fibrosarcoma cell line. The aim of the present study was to evaluate cytotoxicity of D. glabrum root extracts in a human gastric adenocarcinoma (AGS) cell line and explore mechanisms of apoptosis induction, cell cycle arrest and altered gene expression in cancer cells. Materials and Methods: The MTT assay was used to evaluate IC50 values, EB/AO staining to analyze the mode of cell death, and flow cytometry to assess the cell cycle. Quantitative real-time polymerase chain reaction (qRT-PCR) amplification was performed with apoptosis and cell cycle-related gene primers, for cyclin D1, c-myc, survivin, VEGF, Bcl-2, Bax, and caspase-3 to determine alteration of gene expression. Results: Our results showed that n-hexane and chloroform extracts had greatest toxic effects on gastric cancer cells with IC50 values of 6.4 µg/ml and 4.6 µg/ml, respectively, after 72 h. Cell cycle analysis revealed that the population of treated cells in the G1 phase was increased in comparison to controls. Cellular morphological changes indicated induction of apoptosis. In addition, mRNA expression levels of Bax and caspase-3 were increased, and of bcl-2 survivin, VEGF, c-myc and cyclin D1 were decreased. Conclusion: Our study results suggest that D. glabrum has cytotoxic effects on AGS cells, characterized by enhanced apoptosis, reduced cell viability and arrest of cell cycling.

Phytochemical and Antioxidant Investigation of the Aerial Parts of Dorema glabrum Fisch. & C.A. Mey.

Dorema glabrum Fisch. & C.A. Mey. (Apiaceae) is a monocarpic perennial plant distributed in southern Caucasus. In Azerbaijan Republic folk medicine, the gum-resin of this species is used as a diuretic and anti-diarrheal agent. It is also traditionally used for the treatment of bronchitis and catarrh. In the present study, chemical constituents of the essential oil and extract of D. glabrum aerial parts were investigated and their free radical scavenging potentials were assessed. GC-MS and GC-FID analyses of the plant essential oil resulted in identifying twenty compounds, out of which elemicin (38.6%) and myristicin (14.3%) were main compounds. Seven compounds including daucosterol (1), chlorogenic acid (2), a mixture of cynarin (3) and 3,5-di-O-caffeoylquinic acid (4), isorhamnetin-3-O-ß-D-glucopyranoside (5), isoquercetin (6) and astragalin (7) were also isolated from the ethyl acetate and methanol fractions of D. glabrum aerial parts using different chromatographic methods on silica gel (normal and reversed-phase) and sephadex LH20. Structures of the isolated compounds were elucidated using UV and (1)H, (13)C-NMR spectrain comparison with those reported in respective published data. Antioxidant activities of the crude extract, fractions and isolated compounds were evaluated using DPPH free radical scavenging assay method. Among the fractions, methanol fraction (IC50=53.3 ±4.7µg mL(-1)) and among the isolated compounds, caffeoylquinic acid derivatives exhibited the highest free radical scavenging activity (IC50= 2.2-2.6 µg mL(-1)).

Essential oil composition and isolation of freeradical-scavenging phenolic glycosides from the aerial parts of Ajuga chamaepitys growing in Iran

From the methanolic extract of the aerial parts of Ajuga chamaepitys (L.) Schreb., Lamiaceae, one of the Iranian medicinal plants, the phenylethanoid glycoside, acteoside, and two flavone glycosides, chrysoeriol 7-O-glucopyranoside (3'-methoxy-luteolin 7-O-glucopyranoside) and apigenin 7-O-rhamnopyranoside, were isolated by a combination of solid-phase extraction (SPE) and preparative reversed-phase high-performance liquid chromatography (prep-RP-HPLC) methods. Structures of the isolated compounds were elucidated by spectroscopic means. The free-radical-scavenging properties of the extracts, fractions and isolated compounds were determined by the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay. While among the extracts, the MeOH extract showed the highest level of free-radical-scavenging activity (RC50 1.15 × 10-1 mg/mL), chrysoeriol 7-O-glucopyranoside was the most active (RC50 3.00 × 10-3 mg/mL) among the isolated compounds. The GC-MS and the GC-FID analyses revealed α-pinene (23.66%), β-pinene (9.33%), 1-octen-3-ol (9.72%), β-phellandrene (8.70%) and germacrene-D (7.92%) as the major components of the essential oils derived from the aerial parts of this plant. The presence of phenolic glycosides and the α- and β-pinene-rich essential oils in A. chamaepitys may provide some rationale for the traditional medicinal uses of this species in Iran.