Oxadiazole compounds are of great interest because they have a range of biological activities ranging from antioxidants to anticancer agents. Against this background, we wanted to demonstrate the antioxidant, enzyme inhibitory, and anticancer effects of 5(4-hydroxyphenyl)-2-(N-phenylamino)-1,3,4-oxadiazole (Hppo). Antioxidant abilities were measured through free radical scavenging and reducing power tests. Enzyme inhibitory effects were studied by cholinesterases, tyrosinase, amylase, and glucosidase. The anticancer effect was tested on pancreatic cancer cell lines (PANC-1, CRL-169) and on HEK293 cell lines. The compound showed significant antioxidant activity (particularly in the CUPRAC (cupric acid-reducing antioxidant capacity) assay) and enzyme inhibitory properties (particularly glucosidase inhibition). In the anticancer test, the compound showed strong anticancer activity in pancreatic cancer with apoptotic signaling pathways. These results were confirmed by molecular modeling and bioinformatics tools. Thus, our findings can provide novel and versatile compounds for the development of multidirectional drugs in the pharmaceutical industry.

In this study, we conducted a thorough analysis of Rheum turkestanicum (RT) and Calendula officinalis flowers (COF) extracts with varying polarities using LC-MS chemical profiling and biological tests (antioxidant, antimicrobial, enzyme inhibition, and cytotoxic effects). The highest level of total phenolic content in the ethanol extract of RT with 75.82 mg GAE/g, followed by the infusions of RT (65.00 mg GAE/g) and COF (40.99 mg GAE/g). A total of 20 bioactive compounds were identified and quantified. The ethanol extract of COF was rich in terms of 5-O-caffeoylquinic acid (2780.56 μg/g), isorhamnetin-O-rutinoside (1653.59 μg/g), and rutin (1356.97 μg/g). However, RF extracts were rich in catechin gallate (21.66-80.01 μg/g) and 5-O-caffeoylquinic acid. Except for metal chelating ability, the ethanol extract of RT exhibited the strongest ability (DPPH: 171.5 mg TE/g; ABTS: 387.35 mg TE/g; CUPRAC: 449.80 mg TE/g; FRAP: 195.60 mg TE/g; and PBD: 1.52 mmol TE/g). In the enzyme inhibition tests, the tested ethanol extracts for both species were more active than the infusion. The highest values for tyrosinase were recorded as 72.47 mg KAE/g (in RT extracts) and 71.74 mg KAE/g (in COF extracts). Furthermore, all extracts underwent assessment for their antibacterial and antifungal properties, targeting both Gram-positive and Gram-negative bacteria, as well as clinical yeast and fungal microorganisms. In silico studies yielded valuable insights into the potential therapeutic applications of the bioactive compounds identified in COF and RT extracts. Stable interactions were observed between key compounds, such as isorhamnetin 3-O-glucoside and 3-O-caffeoylquinic acid, with crucial target proteins (AChE, BChE, and MurE). These compounds formed stable hydrogen bonds with minimal root mean square deviation (RMSD) fluctuations, particularly in the isorhamnetin 3-O-glucoside-Staphylococcus aureus MurE and 3-O-caffeoylquinic acid-MurE of S. aureus complexes. These findings further underscore the potential of these compounds as promising candidates for therapeutic development.

Agri-food by-products cause significant environmental and annual market losses. This study addresses the challenge of sustainable use of these by-products by developing an innovative food supplement from artichoke residues, rich in active molecules such as inulin and polyphenols. Its key components were identified and quantified using high-performance liquid chromatography with diode-array detection (HPLC-DAD), loaded into zein liposomes for oral delivery, and evaluated in vitro for biological effects. Zein liposomes were characterized by a small size (approximate to 132 nm), homogeneously dispersion (polydispersity index approximate to 0.19), a negative charge (around-44 mV), and an efficient incorporation of artichoke extract (>70%). Freeze-drying improved the stability of vesicle dispersions and small-Angle X-ray Scattering analysis confirmed that zein stabilized the vesicles under acidic conditions. Zein liposomes controlled the release of loaded molecules in gastrointestinal-simulating pH (at pH 1.20 similar to 21% of the chlorogenic acid equivalents were released) and inhibited alpha-amylase (approximate to similar to 0.90 mu g of acarbose equivalents/mL) and alpha-glucosidase (similar to 0.97 mu g of acarbose equivalents/mL) in vitro. Formulations were biocompatible (cell viability approximate to 90%) with Caco-2 cells and protected them from oxidative stress. Overall, the results suggest that zein liposomes loaded with artichoke by-product extract are well-suited as a food supplement or nutraceutical to boost nutritional value.

Sorbus torminalis (L.) Crantz has a rich history of versatile applications spanning the fields of medicine and nutrition. It is noteworthy that the decoction obtained from S. torminalis leaves is a traditional treatment method against both diabetes and stomach disorders. Phytochemical profiling determined by HPLC/MS-MS. The effects of the extracts on cell viability were investigated using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) method against MDA-MB-231 cell line (human breast adenocarcinoma).The ethanol/water extract contained more concentration of total phenolic (91.41 mg gallic acid (GAE) equivalent /gr) and flavanoid (29.10 mg rutin (RE) equivalent/gr) in the tested extract (p<0.05). Resulting of HPLC analysis, the chemical constituents varied depending on the solvents and chlorogenic acid, hyperoside, isoquercetin, delphindin-3,5-diglucoside, procyanidin B2, epicatechin, neochlorogenic acid, 3,5-dicaffeoylquinic acid were identified in all extracts. Overall, ethanol, n-hexane and ethyl acetate extracts showed the highest inhibition for the tyrosinase enzyme. The effect of leaf extracts of S. torminalis on antimicrobial, biofilm inhibitory, and anticancer activities was examined. Based on outcomes of our study recognize this plant as a critical source of medically active chemicals for feasible phytopharmaceutical and nutraceutical applications, providing the first scientific insight into the detailed biological and chemical profiles of S. torminalis.

This study investigates the biochemical composition and biological properties of different parts (leaves, roots, and twigs) of two Cistus species (Cistus monspeliasis and Cistus parviflorus). The extracts were analysed using UHPLC-MS/MS to determine their chemical profiling. A range of antioxidant assays were performed to evaluate the extract’s antioxidant capabilities. The enzyme inhibition studies focused on acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-amylase, and α-glucosidase and tyrosinase. In addition, the study examined the antimicrobial effects on different bacteria and yeasts and evaluated the toxicity using the MTT assay. Quinic acid, citric acid, gallic acid, catechin, quercetin derivatives, kaempferol, myricetin, ellagic acid, prodelphinidins, procyanidins, scopoletin, and flavogallonic acid dilactone are the main bioactive compounds found in both species. In enzyme inhibition assays, C. monspeliasis roots exhibited significant activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), with the values of 2.58 ± 0.02 mg GALAE/g and 11.37 ± 1.93 mg GALAE/g, respectively. Cytotoxicity studies showed mostly weak toxicity, with some samples moderately reducing viability in RAW and HepG2 cells. These findings underscore the diverse biochemical profiles and bioactive potential of Cistus species, suggesting their utility as natural sources of antioxidants and enzyme inhibitors for pharmaceutical and nutraceutical development.

Abstract: Lipidic carriers employed for the entrapment, encapsulation and precise targeting the bioactive compounds offer significant advantages to the biomedical, pharmaceutical, nutraceutical and cosmeceutical industries. These lipid and phospholipid-based vesicles can improve the solubility, permeation, pharmacokinetics, pharmacodynamics and bioavailability of the encapsulated material. As a matter of fact, nano-sized phospholipid vesicles have attained the highest success rate in commercialization among the available drug encapsulation technologies. Nevertheless, developing industrially acceptable products still remains a challenge. Among the main disadvantages of conventional lipidic carriers are their instability in the biological environments, and their sensitivity to changes in the pH, temperature and enzymes. To overcome these problems, the new generation of phospho/lipid-based carrier systems was developed using ingredients from microorganisms living in extremely harsh environments (i.e., archeaeobacteria). The present entry aims to review unique features of archaeolipids and recent developments in their application for the encapsulation of genetic material as well as their role and function as adjuvants and vaccine candidates.

Humans have used medicinal plants to treat various diseases for thousands of years. Cistus species are also widely used in traditional medicine and have various medicinal applications; therefore, they deserve more in-depth research. The present study evaluated the chemical profile, antioxidant, enzyme inhibition, and cytotoxic properties of the twigs and leaves of C. creticus L., C. laurifolius L., and C. salviifolius L. grown in T & uuml;rkiye. The methanolic extracts of the three species were rich in phenolics, mainly flavonoids. Exerted potent antioxidant activity with a methanolic extract from the leaves of C. salviifolius displayed the highest total phenolic (97.08-mg gallic acid equivalent/g) and flavonoid (49.60-mg rutin equivalent/g) contents, as well as antiradical (2,2-diphenyl-1-picrylhydrazyl) assay = 612.11 mg TE (trolox equivalent)/g; (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) = 804.66 mg TE/g), reducing ions (cupric reducing antioxidant capacity) = 690.54 mg TE/g; ferric reducing antioxidant power = 459.34 mg TE/g), and chelating (15.58 mg EDTAE/g) properties. It also revealed the best amylase and glucosidase inhibitory activity. Extracts from the twigs of the three Cistus species, except the leaves of C. salviifolius and C. laurifolius, displayed comparable acetylcholinesterase inhibitory activity (2.48-2.57 mg galanthamine equivalent (GALAE)/g). The twig of C. laurifolius also exerted the best antibutyrylcholinesterase (10.50 mg GALAE/g) and antityrosinase (73.15 mg kojic acid equivalent/g) activities. C. creticus leaves revealed toxicity toward the RAW cell line (cell viability reduced to 68.8%) and were not toxic to normal cells (S17). In conclusion, these three Cistus species were shown to be a rich source of bioactive compounds with the potential for future applications in the food, pharmaceutical, and cosmetic industries.

Stellaria media L., also called chickweed, is widespread in all parts of the world. In the present study, we investigated the biological properties and chemical profiles of different extracts (ethyl acetate, ethanol, ethanol/water, and water) of S. media. The chemical profiles were examined using UHPLC/MS/MS technique. Regarding the biological properties, antioxidant properties as well as enzyme-inhibiting and cytotoxic effects of the extracts were demonstrated by in vitro methods. To obtain further information about the structure-ability relationship, network pharmacology and molecular docking were also performed. Twelve phenolic compounds were identified in the extracts and most of them were flavonoids (apigenin, kaempferol derivatives, etc.). The water extract showed the best free radical scavenging activity, while the ethanol was the most active in reducing power tests. When inhibiting AChE, the ethyl acetate extract showed the best inhibitory effect. The water extract has a good cytotoxic effect on HepG2 (cell viability: 33.9% at a concentration of 100 g/mL). The analysis, performed using the STRING database, included these 45 cancer-associated targets. The identified hub genes were TP53, CDKN2A, PTEN, KRAS, and HRAS. In molecular docking analysis, acacetin-O-hexoside-O-deoxyhexoside and napigenin-7-O-hexoside exhibit remarkable binding energies with proteins. Consequently, S. media can be potential raw materials for designing functional formulations in the pharmaceutical, nutraceutical, and cosmeceutical industries.

Ocimum species have a great interest in different traditional medicinal systems. This study examined the chemical composition, antioxidant properties, enzyme inhibitory effects, and antibacterial and antifungal activities of the aerial parts of Ocimum gratissimum, Ocimum americanum, and Ocimum basilicum from the Comoros Islands. The extracts were analyzed using high-performance liquid chromatography-mass spectrometry (HPLC-MS) to determine their chemical composition. Antioxidant activity was assessed using 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), cupric reducing antioxidant capacity (CUPRAC), ferric reducing antioxidant power (FRAP), chelating ability, and phosphomolybdenum radical scavenging assays. Enzyme inhibitory activities against acetylcholinesterase (AChE), butrylcholinesterase (BChE), tyrosinase, amylase, and glucosidase were evaluated using spectrophotometric methods. Antibacterial and antifungal activities were tested using the broth microdilution method against selected pathogenic microorganisms. The selected enzymes and proteins were evaluated using in silico methods with biomolecules from these plants. In addition, 111 different metabolites were identified in the tested extracts using advanced HPLC/MS techniques. The most significant number of detected compounds were derivatives of hydroxycinnamic acids, followed by flavonoid glycosides and aglycones and derivatives of hydroxybenzoic acids. All three Ocimum species exhibited significant antioxidant activities, O. gratissimum exhibited the best-reducing abilities in CUPRAC and FRAP assays. In addition, enzyme inhibitory assays revealed that O. americanum had the most potent inhibitory effect on tyrosinase (48.01 ± 3.89 mg kojic acid equivalent [KAE]/g), and amylase (1.08 ± 0.02 mmol acarbose equivalent [ACAE]/g). Antibacterial and antifungal tests demonstrated that the extracts possess broad-spectrum activity. Molecular docking results showed that compounds exhibited remarkable binding energies with target enzymes and proteins. The molecular dynamics simulations identified chicoric acid with MurE of Staphylococcus aureus complex as the most promising drug candidate. These findings support their traditional medical and nutraceutical uses and suggest possibilities for natural functional applications.

The members of the genus Centaurea have a great interest in pharmaceutical and nutraceutical fields due to their biological potential. Based on this information, we aimed to evaluate the biological properties (antioxidant, enzyme inhibition and cytotoxicity) and chemical profile of the extract of Centaurea stapfiana, , an unstudied species. The highest total phenolic content was found in the ethanol/water extract with 32.17 mg GAE/g. A total of 102 of them were identified by HPLC-ESI-QTOF-MS analysis. These compounds were mainly hydroxybenzoic acid and hydroxycinnamic acid as well as flavonoids. In the antioxidant tests, the ethanol/water extract had the best free radical scavenging and reducing ability. However, in the enzyme inhibition test, the ethanol extract was the most active. The extracts were also tested on two tumour cell lines (RAW 264.7 and HepG2) and one non- tumour cell line (S17). The ethanol extract showed the promising effect on HepG2 (cell viability: 28.6 % at 50 g/ml). Furthermore, we examined the interactions between the compounds and enzymatic and cellular targets. A good interaction was found between quercetin-3-xylosyl-(1- > 6)-glucoside and iNOS. In summary, our results suggest that C. stapfiana can be considered as a versatile raw material for the development of health- promoting applications in the pharmaceutical and cosmeceutical fields.

The members of the genus Marrubium are of great interest because they contain biologically active compounds. With in this mind, we aimed to examine the chemical profiles and biological activities (antioxidant, enzyme inhibition and antimicrobial effects) of different extracts of Marrubium astracanicum subsp. macrodon. Extractions of the plants were carried out with the solvent n-hexane, acetone, acetone/water (70 %), and water. The antioxidant activity of these extracts was evaluated through six different assays, namely 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2 '-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), cupric reducing antioxidant capacity (CUPRAC), ferric reducing antioxidant power (FRAP), metal chelating ability (MCA), and phosphomolybdenum (PBD). Additionally, the inhibitory effects of these solvents on five enzymes acetylcholinesterase (AChE), butyrylcholinesterase (BChE), tyrosinase, amylase, and glucosidase were investigated. The study detected 38 compounds in a plant, with the highest content in acetone and acetone/water extracts. The acetone/water extract demonstrated the highest antioxidant activity aligning with its superior total phenolic (51.9 mg gallic acid equivalent (GAE)/g) and flavonoid (31.0 mg rutin equivalent (RE)/g) content, highlighting a direct correlation between bioactive compound concentration and antioxidant potential across various assays. Regarding the enzyme inhibition activity, the acetone/water extract of this plant exhibited significant AChE inhibition measuring 2.5 mg galantamine equivalent (GALAE)/g and anti-glucosidase activity measuring 1.2 mmol acarbose equivalent (ACAE)/g. In antimicrobial evaluation, the acetone extract exhibited good antimicrobial activity against B. cereus at 0.2 mg/ml dose. In molecular docking analysis, delphinidin 3,5 diglucoside showed a good interaction with the active site of AChE active. Through the application of network pharmacology, an in-depth exploration was conducted to unravel the potential of compounds from M. astracanicum against Alzheimer's disease. The study specifically focused on identifying key targets within the Alzheimer's disease pathway, pinpointing crucial proteins such as Glycogen synthase kinase-3 (GSK3), Prostaglandin-endoperoxide synthase 2 (PTGS2), and Alzheimer's-associated amyloid precursor protein (APP). In summary, the plant can serve as a valuable ingredient in the production of functional products within the pharmaceutical, nutraceutical, and cosmeceutical applications.

The genus Paeonia has gained great interest from the scientific community for exploration as a source of bioactive compounds. The current work focused on the chemical characterisation and biological properties of Paeonia mascula L. from Turkey. The chemical characterisation of the extracts was analysed by LC-DAD-ESI and LC-APCI-MS. The antioxidant, enzyme inhibitory and anticancer properties were evaluated in vitro. The methanol extract was the most effective as antioxidant, metal chelator and was the most effective inhibitor of acetylcholinesterase (AChE) and tyrosinase. The chloroform extract showed highest inhibition of butrylcholinesterase (BChE) and amylase. Ethyl acetate extract displayed glucosidase inhibition. The most abundant compounds were hexagalloyl glucose, penta galloyl glucose, gallic acid, tetragalloyl glucose, paeoniflorin and the most abundant flavonoid was quercetin-3-O-glucoside. β-sitosterol was the most abundant phytosterol. The anticancer effect of P. mascula was evaluated by using HELA cells evaluating different pathways. The possible inhibition effect of P. mascula on the reactive oxygen species (ROS) pathway was controlled by performing the advanced glycation end product (AGE). To determine the effect of P. mascula on active molecular pathways in the HELA cancer cell line, the levels of Phospho-NF-Kβ p65, AGE receptor RAGE protein, antiapoptotic BCL-2, proapoptotic BAX and P-53 proteins were checked, respectively. Phospho-NF-Kβ p65, RAGE and BCL-2, which are both ROS inducers and play an active role in cancer prognosis, were found to have decreased levels after the application of P. mascula. Our findings provide a scientific basis for Paeonia mascula, which may serve as a source of naturally occurring bioactive compounds for healthpromoting applications.

Arum elongatum (Araceae) is widely used traditionally for the treatment of abdominal pain, arterial hypertension, diabetes mellitus, rheumatism and hemorrhoids. This study investigated the antioxidant properties, individual phenolic compounds, total phenolic and total flavonoid contents (HPLC/MS analysis), reducing power and metal chelating effects of four extracts obtained from A. elongatum (ethyl acetate (EA), methanol (MeOH), methanol/water (MeOH/water) and infusion). The inhibitory activity of the extracts were also determined against acetylcholinesterase, butyrylcholinesterase, tyrosinase, amylase and glucosidase enzymes. The MeOH/water extracts contained the highest amount of phenolic contents (28.85 mg GAE/g) while the highest total flavonoid content was obtained with MeOH extract (36.77 mg RE/g). MeOH/water demonstrated highest antioxidant activity against DPPH⋅ radical at 38.90 mg Trolox equivalent per gram. The infusion extract was the most active against ABTS+ ⋅ (133.08 mg TE/g). MeOH/water extract showed the highest reducing abilities with the CUPRAC value of 102.22 mg TE/g and the FRAP value of 68.50 mg TE/g. A strong metal chelating effect was observed with MeOH/water extract (35.72 mg EDTAE/g). The PBD values of the extracts ranged from 1.01 to 2.17 mmol TE/g. EA extract displayed the highest inhibitory activity against AChE (2.32 mg GALAE/g), BChE (3.80 mg GALAE/g), α-amylase (0.56 mmol ACAE/g) and α-glucosidase (9.16 mmol ACAE/g) enzymes. Infusion extract was the most active against tyrosinase enzyme with a value of 83.33 mg KAE/g. A total of 28 compounds were identified from the different extracts. The compounds present in the highest concentration were chlorogenic acids, 4-hydroxybenzoic acid, caffeic acid, p-coumaric acid, ferulic acid, isoquercitrin, delphindin 3,5-diglucoside, kaempferol-3-glucoside and hyperoside. The biological activities of A. elongatum extracts could be due to the presence of compounds such as gallic acid, chlorogenic acids, ellagic acid, epicatechin, catechin, kaempferol, 4-hydroxybenzoic acid, caffeic acid, p-coumaric acid, ferulic acid, quercetin, isoquercitrin, and hyperoside. Extracts of A. elongatum showed promising biological activities which warrants further investigations in an endeavor to develop biopharmaceuticals.

Several species within the genera Cassia or Senna have a treasure of traditional medicines worldwide and can be a promising source of bioactive molecules. The objective of the present study was to evaluate the phenolic content and antioxidant and enzyme inhibition activities of leaf methanolic extracts of C. fistula L., C. grandis L., S. alexandrina Mill., and S. italica Mill. The two Cassia spp. contained higher total polyphenolic content (42.23–49.75 mg GAE/g) than the two Senna spp., and C. fistula had significantly (p ˂ 0.05) the highest concentration. On the other hand, the Senna spp. showed higher total flavonoid content (41.47–59.24 mg rutin equivalent per g of extract) than that found in the two Cassia spp., and S. alexandrina significantly (p ˂ 0.05) accumulated the highest amount. HPLC–MS/MS analysis of 38 selected bioactive compounds showed that the majority of compounds were identified in the four species, but with sharp variations in their concentrations. C. fistula was dominated by epicatechin (8928.75 µg/g), C. grandis by kaempferol-3-glucoside (47,360.04 µg/g), while rutin was the major compound in S. italica (17,285.02 µg/g) and S. alexandrina (6381.85). The methanolic extracts of the two Cassia species exerted significantly (p ˂ 0.05) higher antiradical activity, metal reducing capacity, and total antioxidant activity than that recorded from the two Senna species’ methanolic extracts, and C. fistula displayed significantly (p ˂ 0.05) the highest values. C. grandis significantly (p ˂ 0.05) exhibited the highest metal chelating power. The results of the enzyme inhibition activity showed that the four species possessed anti-AChE activity, and the highest value, but not significantly (p ≥ 0.05) different from those obtained by the two Cassia spp., was exerted by S. alexandrina. The Cassia spp. exhibited significantly (p ˂ 0.05) higher anti-BChE and anti-Tyr properties than the Senna spp., and C. grandise revealed significantly (p ˂ 0.05) the highest values. C. grandise revealed significantly (p ˂ 0.05) the highest α- amylase inhibition, while the four species had more or less the same effect against the α-glucosidase enzyme. Multivariate analysis and in silico studies showed that many of the identified phenols may play key roles as antioxidant and enzyme inhibitory properties. Thus, these Cassia and Senna species could be a promising source of natural bioactive agents with beneficial effects for human health.

The genus Scorzonera is one of the most important genera in the Asteraceae family, and members of the genus have great potential in traditional medicine systems. In this article we aimed to determine skin protective and antimicrobial effects of different extracts from the roots and aerial parts of three Scorzonera species (S. hieraciifolia, S. hispanica, S. tomentosa). Antimicrobial properties were evaluated against different pathogen bacterial and fungal strains, whereas protective effects were studied in isolated mouse skin specimens exposed to hydrogen peroxide. In this biological model, the levels of l-dopa and prostaglandin E-2 (PGE(2)) were evaluated, as markers of tyrosinase activity and inflammation, respectively. All extracts were effective in blunting the increased hydrogen peroxide-induced l-dopa turnover and PGE(2) level; thus, suggesting skin protection that could be partly related to the content of flavonoids, among which rutin, in the tested extracts. Additionally, all extracts were effective as antibacterial and antifungal agents; intriguing results were observed against Trichoderma viride, which was particularly sensitive to S. tomentosa root extracts. Therefore, the present study strengthens the importance of investigating herbal preparations obtained from plants belonging to the genus Scorzonera, for the management and prevention of inflammatory and infectious skin conditions.

In the present study, extracts of four medicinal plants (Cajanus cajan, Carica papaya, Morinda morindoides, and Senna alexandrina) obtained at two different extraction temperatures (24 and 45 degrees C) were investigated for their polyphenolic contents, antioxidant, and enzyme inhibitory properties. For instance, extracts of the studied species yielded total phenolic and flavonoid contents in the range of 17.8-66.3 mg GAE/g and 4.18-26.8 mg RE/g, respectively. High performance liquid chromatography (HPLC) coupled with antioxidant assays (HPLC-FRAP and HPLC-DPPH) was employed for direct identification of antioxidant compounds. The extracts showed moderate to high antioxidant potentials in various assays performed (radical scavenging activity: 10.15-240.08 mg TE/g; reducing activity: 24.88-229.59 mg TE/g; metal chelating activity: 5.56-18.32 mg EDTAE/g and total antioxidant capacity: 1.40-2.18 mM TE/g). Additionally, the extracts were found to exert inhibitory properties against all the studied enzymes (anti-acetylcholinesterase: 1.80-2.29 mg GALAE/g; anti-butyrylcholinesterase: 0.19-3.13 mg GALAE/g; anti-amylase: 0.39-0.58 mmol ACAE/g; anti-glucosidase: 0.17-1.36 mmol ACAE/g and anti-tyrosinase: 60.97-73.71 mg KAE/g). Of all, C. cajan extracts were revealed to be the best antioxidants and inhibitors of most enzymes. Therefore, this study demonstrated the pharmacological potentials of the four medicinal plants, especially C. cajan as viable sources of phyto-ingredients that could be of interest in nutraceutical development. Novelty impact statement The methanol extracts from four medicinal plant with different extraction temperatures (24 and 45 degrees C) were investigated. HPLC-DPPH and HPLC-FRAP methodologies were used. Different levels of phenolics in the tested extracts were determined. Cajamus cajan extracts were found to be the best one in the antioxidant assays. Our findings could be opened valuable windows in the further pharmaceutical studies and applications

Mentha spicata is one of the most popular species in the genus, and it is of great interest as a gastrointestinal and sedative agent in the folk medicine system. In this study, different M. spicata extracts, obtained by the use of four solvents (hexane, chloroform, acetone and acetone/water) were chemically characterized using HPLC-ESI-MS n, which allowed for identification of 27 phenolic compounds. The extracts’ antioxidant and enzyme inhibitory properties were investigated. In addition, neuroprotective effects were evaluated in hypothalamic HypoE22 cells, and the ability of the extracts to prevent the hydrogen peroxide-induced degradation of dopamine and serotonin was observed. The best antioxidant effect was achieved for all the extraction methods using acetone/water as a solvent. These extracts were the richest in acacetin, eriodictyol, hesperidin, sagerinic acid, naringenin, luteolin, chlorogenic acid, chrysoeriol and apigenin. The intrinsic antioxidant and enzyme inhibition properties of the acetone/water extract could also explain, albeit partially, its efficacy in preventing prostaglandin E2 overproduction and dopamine depletion (82.9% turnover reduction) in HypoE22 cells exposed to hydrogen peroxide. Thus, our observations can provide a scientific confirmation of the neuromodulatory and neuroprotective effects of M. spicata.

Withania somnifera (L.) Dunal has been identified as a miracle herb, thus the present study was carried out to investigate W. somnifera growing in Cholistan Desert for its secondary metabolic profile and various biological activities. The WS-C fraction of methanolic extract (WS-M) of W. somnifera was found rich in phenolics (39.96 +/- 0.90 mg GAE/g extract), and it exhibited significant DPPH (30.69 +/- 0.78 mg TE/g extract) and ABTS (113.60 +/- 2.41 mg TE/g extract) free radical inhibitory activities, cupric (Cu+2) and ferric (Fe+3) reducing potential as 157.51 +/- 2.10 and 124.60 +/- 0.34 mg TE/g extract. The same fraction also exhibited strong total antioxidant capacity (1.39 & PLUSMN; 0.01 mmol TE/g extract) in phosphomolybdenum assay. Other fractions displayed relatively lower potential. In metal-chelating assay, WS-M, WS-E and WS-B showed nearly equal potential with values of 22.35 +/- 0.62, 20.96 +/- 0.12 and 22.10 +/- 0.75 mg EDTAE/g of the extract, while in AChE, BChE and tyrosinase inhibitory assay, again WS-C fraction was most active (7.50 +/- 0.21, 8.01 +/- 0.53 mg GALAE/g extract and 36.81 +/- 0.20 mg KAE/g extract, respectively). In alpha-amylase and alpha-glucosidase inhibitory assay, WS-C showed significant potential with values as 0.74 +/- 0.01 and 1.14 +/- 0.004 mmol ACAE/g extract, respectively. UHPLC-MS analysis of WS-C fraction leads to identify 100 secondary metabolites of phenolic, withanolide, terpenoid, ste-roid, lignin, flavonoid and limonoid classes of compounds. This profile makes W. somnifera a fascinating plant. These findings were validated by computational studies, which revealed that selected compounds exhibited high binding free energy and inhibition constants with the enzymes tested. Pearson correlation analysis clearly established that the phenols in the tested extracts were the main players as antioxidants and enzyme inhibitors, which was also substantiated through Principal Component Analysis (PCA). These results suggest that chloro-form might be suitable for preparing further applications with W. somnifera extracts, and this plant can be placed in the list of top-selling herbs, and may become source of commercial assets.

In this study, the methanolic and infusion extracts of two species, Thymbra capitata and Thymus sipyleus subsp. rosulans, were tested for their chemical composition and biological abilities (antioxidant, enzyme inhibitory and anti-inflammatory effects). The extracts yielded total phenolic and flavonoid contents in the range of 83.43–127.52 mg GAE/g and 9.41–46.34 mg RE/g, respectively. HPLC analysis revealed rosmarinic acid to be a major component of the studied extracts (15.85–26.43%). The best ABTS radical scavenging ability was observed in the methanol extract of T. capitata with 379.11 mg TE/g, followed by in the methanol extract of T. sipylus (360.93 mg TE/g). In the CUPRAC assay, the highest reducing ability was also found in the methanol extract of T. capitata with 802.22 mg TE/g. The phosphomolybdenum ability ranged from 2.39 to 3.61 mmol TE/g. In terms of tyrosinase inhibitory effects, the tested methanol extracts (83.18–89.66 mg KAE/g) were higher than the tested water extracts (18.74–19.11 mg KAE/g). Regarding the BChE inhibitory effects, the methanol extracts were active on the enzyme while the water extracts showed no inhibitory effect on it. Overall, the methanolic extracts showed better enzyme inhibition compared to the infusion extracts. Molecular docking also showed the selected exhibited potential binding affinities with all enzymes, with a preference for cholinesterases. Additionally, the extracts were effective in attenuating the LPS-induced increase in COX-2 and IL-6 gene expression in isolated colon, thus indicating promising anti-inflammatory effects. The preliminary results of this study suggest that these species are good natural sources of antioxidants and also provide some scope as enzyme inhibitors, most likely due to their bioactive contents such as phenolic acids, and thus can be exploited for different applications related to health promotion and disease prevention.

Calendula officinalis L., commonly known as pot marigold, is a popular ornamental and highly medicinal plant. In fact, it is well recognized for its wide-ranging therapeutic effects conducive to wound healing and for treating various illnesses. In this study, the essential oils from flowers and leaves of C. officinalis were analysed for their chemical constituents and screened for their in vitro antioxidant and enzyme inhibitory properties. Thirty compounds were found in common in both essential oils, with the sesquiterpene alcohol, alpha-cadinol as the most abundant compound (leaf: 32.3% and flower: 31.3%, respectively). Besides, the essential oils were found to possess antioxidant capacity through free radical scavenging and reducing mechanisms (DPPH: 2.94, 1.52 mg trolox equivalent (TE)/g of oil; ABTS: 20.72, 2.86 mg TE/g; CUPRAC: 31.16, 47.98 mg TE/g; FRAP: 24.84, 23.42 mg TE/g; phosphomolybdenum: 4.01, 4.68 mmol TE/g for leaves and flowers essential oils, respectively). Moreover, the essential oils (leaves and flowers, respectively) were found to inhibit all the tested enzymes, namely amylase (0.46, 0.42 mmol acarbose equivalent (ACAE)/g), glucosidase (1.35, 1.31 mmol ACAE/g), acetylcholinesterase (2.48, 2.36 mg galantamine (GALAE)/g) and butyrylcholinesterase (8.13, 8.20 mg GALAE/g) and tyrosinase (47.73, 54.36 mg kojic acid equivalent/g). Molecular docking was also performed revealing the compounds neophytadiene and alpha-humulene to show the highest binding affinity with tyrosinase. Therefore, the obtained results demonstrated some appealing activities of C officinalis essential oils as natural pharmacological agents that could be valuable for the potential development of phyto-formulations against the herein targeted diseases and especially for cosmeceutical applications.