Bromophenol derivatives were evaluated for their inhibitory effects on three key enzymes associated with diabetes: aldose reductase (AR), α-glycosidase, and α-amylase. In vitro enzyme inhibition assays revealed that Compound 6 exhibited the strongest α-glycosidase inhibition (IC50 = 11.948nM), while Compound 5 showed the highest potency against AR (IC50 = 1.386µM) and α-amylase (IC50 = 5.588nM). Selected compounds (5, 6, 7, and 10) displayed low cytotoxicity against normal human fibroblast cells, indicating favorable safety profiles. Complementary induced-fit molecular docking analyses supported the experimental results, highlighting specific interactions between the compounds and the enzyme active sites. These findings demonstrate that bromophenol derivatives possess significant enzyme-inhibitory potential and may serve as promising multifunctional agents for diabetes management. Induced-fit docking studies revealed that Compounds 6, 10, 5, and 7 exhibited the most favorable binding energies across the tested enzymes, consistent with their high inhibitory potential. Compounds 6 and 10 showed strong α-glucosidase binding through key interactions such as π-π stacking with TRP539/PHE575 and TRP406, as well as extensive hydrogen and halogen bonding with catalytic residues including TYR299, ASP203, ASP327, and ARG526. Similarly, Compounds 5 and 7 formed stable docking poses within α-amylase and aldose reductase via multiple hydrogen bonds, halogen contacts, and π-π stacking with crucial residues like ASP197, GLH233, ASN160, TRP20, and TRP219, highlighting their strong affinity and target-specific interactions.

Taraxacum mirabile Wagenitz, one of the endemic riches of Anatolia, is a species that has remained largely unexplored regarding its enzyme inhibition profile despite its pharmacological potential. The effects of T. mirabile aerial and root extracts, obtained at different polarities, were scrutinized in this study against two important enzymes: lactoperoxidase (LPO), which plays a vital role in the innate immune system, and xanthine oxidase (XO), which is prominently associated with hyperuricemia and oxidative stress. The aerial and root portions of the plant were extracted into fractions of varying polarities using petroleum ether, dichloromethane, ethyl acetate, and butanol. LPO was isolated from buffalo milk (881.6-fold purification, 22.5% yield, and 1249.9 EU/mg specific activity) via affinity chromatography and used in in vitro inhibition assays alongside commercial bovine XO enzyme. The results showed that the ethyl acetate fraction of the aerial part of the plant exhibited the strongest LPO inhibition (IC50: 15.60 ± 0.77 µg/mL) among the fractions. The petroleum ether fraction of both the aerial part (IC50: 11.17 ± 0.94 µg/mL) and the root part (IC50: 11.61 ± 0.59 µg/mL) had the highest inhibitory effect for the XO enzyme. These distinct inhibition profiles allow for significant insights into how plant extracts with varying polarities modulate XO and LPO enzymes. In conclusion, the significant inhibitory activity of T. mirabile extracts toward LPO and XO enzymes highlights their potential as a natural source for developing effective enzyme inhibitors, which could be useful for therapeutic applications.

ABSTRACT The biological potential of a series of primary carbamate derivatives was investigated, focusing on their inhibitory effects against two clinically significant human carbonic anhydrase isoenzymes, hCA I and hCA II. These enzymes are closely associated with various pathological conditions, including glaucoma, epilepsy, and certain types of cancer. The carbamate compounds, previously synthesized via a safer and more sustainable one‐pot method from alcohols, were evaluated for their enzyme inhibition profiles as well as their antioxidant, antibacterial, and antifungal activities. The findings highlight key structure–activity relationships, particularly emphasizing the role of aromatic and electron‐donating substituents in enhancing biological efficacy. This work contributes to the development of multifunctional small molecules with therapeutic relevance. Among the tested compounds, 3 g and 3f , featuring benzyl and methyl substituents, respectively, exhibited the highest inhibitory activities, with 3 g being the most potent. The antioxidant properties of the carbamates were assessed using DPPH radical scavenging, ABTS cation radical scavenging, and Fe 3 + reducing assays. Compound 3 g demonstrated the strongest antioxidant activity, which can be attributed to the electron‐donating effects of its benzyl and methyl substituents. Antimicrobial evaluations against a range of bacterial and fungal strains further revealed that compound 3 g possessed the most potent antibacterial and antifungal activities. Additionally, molecular docking studies were performed to provide complementary insights and support the experimental findings on the biological activities of the carbamate derivatives. Overall, this research underscores the potential of carbamate derivatives as multifunctional bioactive compounds with promising enzyme inhibition, antioxidant, and antimicrobial properties.

This study aimed to investigate the phytochemical composition, antioxidant activity, and enzyme inhibition profile of Citrullus colocynthis and its synthesised silver nanoparticles (AgNPs). Silver nanoparticles (AgNPs) were synthesised using C. colocynthis fruit extract and characterised using various techniques, including UV-Visible Spectroscopy, SEM, XRD, and FTIR, which confirmed the successful formation of AgNPs. The AgNPs exhibited significant antioxidant activity, with IC50 values of 0.45 ± 0.4 µg/mL, 0.45 ± 0.5 µg/mL, and 0.45 ± 0.1 µg/mL for DPPH, FICA, and HRSC assays, respectively. The nanoparticles also showed potent enzyme inhibition activity against urease, carbonic anhydrase, and xanthine oxidase, with IC50 values of 90 ± 0.1 μg/mL, 221 ± 0.4 µg/mL, and 187 ± 0.9 µg/mL, respectively. Notably, the AgNPs demonstrated enhanced bioactivity compared to the plant extract, highlighting their potential as therapeutic agents. This study provides new insights into the antioxidant and enzyme inhibition potential of C. colocynthis AgNPs, warranting further investigation into their therapeutic applications.

Abstract Plant‐derived phenolic and flavonoid compounds exhibit significant health benefits, encompassing antioxidant and enzyme inhibitory activities. We compared three extraction techniques—maceration (MAC‐ME), Soxhlet extraction (SOE‐ME), and ultrasound‐assisted extraction (UAE‐ME)—for their effectiveness in recovering bioactive compounds from Anthemis pseudocotula and assessed the resulting antioxidant and enzyme inhibition profiles. MAC‐ME and UAE‐ME produced higher total phenolic contents than SOE‐ME, with MAC‐ME achieving the greatest yield. In antioxidant assays, MAC‐ME showed superior performance in the phosphomolybdenum and ferrous ion chelating tests. Conversely, UAE‐ME delivered the strongest reducing power in the CUPRAC and FRAP assays, and both MAC‐ME and UAE‐ME outperformed SOE‐ME in ABTS radical scavenging. Regarding enzyme inhibition, MAC‐ME and UAE‐ME exhibited more potent acetylcholinesterase (AChE) inhibition than SOE‐ME; SOE‐ME was most effective against tyrosinase, whereas MAC‐ME showed the highest α‐amylase inhibition. Overall, MAC‐ME proved the most effective for extracting phenolic and flavonoid compounds from A. pseudocotula , with UAE‐ME also demonstrating notable antioxidant efficacy, underscoring the impact of extraction technique on bioactivity.

ABSTRACTSince plant essential oil contains medicinally valuable compounds, its usability as a pharmacotherapeutic agent has been the focus of attention within the century's needs. The lack of sufficient studies on the medical and pharmacological evaluation of Fibigia clypeata (L.) Medik has made this plant the target of our study. This study analyzes the phytochemical composition and biological activity of F. clypeata essential oil. As a result, dimethyl disulfide and dimethyl trisulfide were found to be the main compounds of the plant essential oil, with rates of 73.13% and 19.87%, respectively. The antifungal property of plant essential oil is more effective than its antibacterial property, with MIC values ranging between 0.039 and 0.312 μL/mL for fungal species and up to 3.750 μL/mL for bacterial species. The enzyme inhibition profiles were investigated towards two enzymes, namely, anticholinesterase and α‐glucosidase, targeted for anti‐diabetic studies. Anticholinesterase activity was proved with the IC50 values of 17.31 and 4.78 μg/mL for Acetylcholinesterase (AChE) and Butyrylcholinesterase (BChE) enzymes, respectively. DPPH and CUPRAC activities were the most promising antioxidant studies, with values of 1.54 and 3.72 μg/mL. It was observed that α‐Terpineol made a hydrogen bond with ASN80, and 1‐(2,6,6‐Trimethyl‐1,3‐cyclohexadien‐1‐yl)ethanol made a hydrogen bond with SER82. Although molecular dock scores were better for antifungal activity, it was determined that no interactions, such as hydrogen bonding or pi interaction, were observed. This preliminary study showed that F. clypeata essential oil is a natural source with promising in vitro antimicrobial, antioxidant, and anticholinesterase activities that warrants further investigation, including safety assessments, due to the high concentration of sulfur‐containing compounds. Molecular docking and ADME prediction results showed that α‐Terpineol and 1‐(2,6,6‐Trimethyl‐1,3‐cyclohexadien‐1‐yl)ethanol were more prone to antimicrobial activity.

Harnessing the bioactive potential of the Algerian endemic Albuca amoena (Batt.) J.C. Manning and Goldblatt (Asparagaceae): new insights into antioxidant activity, enzyme inhibition, and phytochemical profile

Determination of Antioxidant Activity, Vitamin C Content, GST Enzyme Inhibition and Phenolic Profile by LC-HRMS of Jujube Fruit and Seed

This study focuses on the synthesis and characterization of novel sitagliptin derivatives, aiming to develop potent, orally active anti-diabetic agents with minimal side effects for the management of type 2 diabetes mellitus. Copper (II) (SCu1-SCu9) and zinc (II) (SZn1-SZn9) metal complexes of sitagliptin-based derivatives were synthesized via a template reaction. The synthesized complexes were comprehensively characterized using elemental analysis, FTIR, UV-Vis, 1 h NMR, and 13C NMR spectroscopy. The biological efficacy of these compounds was assessed through α-amylase and α-glucosidase enzyme inhibition assays, with molecular simulation studies providing additional confirmation of their inhibitory activity. Among the tested derivatives, SD7, SD4, SD3, SD5, and SD9 demonstrated enzyme inhibition profiles comparable to the standard inhibitors. However, the metal complexes exhibited absorption challenges, which may influence their bioavailability. These findings highlight the significant anti-diabetic potential of the synthesized compounds against targeted enzymes, establishing a foundation for their development as lead molecules in future therapeutic research.

Nizip Yaglık olive leaves (Olea europaea L.) collected at different seasons and altitudes: Enzyme inhibition, antioxidant activities and phenolic compound profiles

Justicia vahliiRoth. is an important wild medicinal food plant traditionally used for treating inflammation and various common ailments. This study investigated the chemical composition, antioxidant, enzyme inhibition and toxicity profiles of n-hexane (nHEJv) and chloroform (CEJv) extracts of J. vahlii. Moreover, the effect of the extracts was evaluated on CCl4 induced liver injury. The total phenolic and flavonoid contents were present in both extracts in significant amount. The UPLC-Q-TOF-MS and GC-MS profiling of CEJv tentatively identified several important phytocompounds. The CEJv extract was comparatively more active for antioxidant activity and α-amylase inhibition, whereas the nHEJv extract presented higher inhibition potential against urease, tyrosinase, and α-glucosidase enzymes. Similarly, the in-silicostudy of four major compounds, i. e., 1-acetoxypinoresinol, 3-hydroxysebacic acid, nortrachelogenin, and viscidulin-III have shown a good docking score against the clinically significant enzymes. The acute oral toxicity and brine shrimp lethality assaysrevealed the extracts as non-toxic. The CCl4 treated animals showed a geared depletion of various antioxidant enzymes which were significantly reversed with the treatment of the extracts. Overall, the study's findings revealed J. vahliiwith antioxidant mediated hepatoprotective and enzyme inhibition potential and warrant further research on isolation of the bioactive compounds.

Heavy metals altered the xenobiotic metabolism of rats by targeting the GST enzyme: An in vitro and in silico study

Interest in the use of bryophytes in pharmaceutical, cosmetic, and food industrial applications is growing worldwide due to their secondary metabolites. In this study, n-hexane crude extracts and further fractions (aqueous, ethyl acetate and n-butanol) of aqueous ethanol (80:20, ethanol:H2O, v/v) were obtained from five different bryophytes (Pellia epiphylla, Conocephalum conicum, Porella platyphylla, Plagiomnium cuspidatum and Mnium spinulosum) collected from Trabzon, Türkiye. The total phenolic compound (TPC) content, antioxidant capacity (AC) and enzyme inhibition activity (acetylcholine esterase, butyrylcholine esterase, tyrosinase, α-amylase and α-glucosidase) of the extracts and fractions were species-specific and varied significantly between the crude extracts and fractions. Among the different bryophytes, Porella platyphylla and Pellia epiphylla in n-butanol and Plagiomnium cuspidatum and Mnium spinulosum in ethyl acetate fraction exhibited the highest TPC contents and AC values. The contents of phenolic acids liberated in free, ester and glycoside forms were also species-specific. p-Hydroxybenzoic acid (p-HBA) in free form in P. cuspidatum and P. platyphylla, p-coumaric acid (p-CoA) in ester form and m-hydroxybenzoic acid (m-HBA) in glycoside form in M. spinulosum were the major phenolic acids in the bryophytes. The n-hexane extracts of the bryophytes, in particular M. spinulosum, had IC50 values ​​almost 100 times lower than acarbose. This suggests that M. spinulosum in particular may represent a possible candidate for the production of new antidiabetic agents.

Novel Mitochondrial and DNA Damaging Fluorescent Calix[4]arenes Bearing Isatin Groups as Aromatase Inhibitors: Design, Synthesis and Anticancer Activity

Objective: In today's world where chronic diseases are increasing, research on effective treatment methods continues.Different methods and drug development studies for treatment continue in a multidisciplinary manner.In order to reduce side effects, natural products are the primary choice and are in the first place in research. Materials and Methods:The present study was undertaken to study enzyme-inhibiting activity of ethyl acetate and methanol extracts from Trifolium arvense and Medicago lupulina in order to explore these plants for their therapeutic use.Inhibition effects for cholinesterases (acetylcholinesterase and butyrylcholinesterase), tyrosinase, -amylase and -glucosidace tests were performed. Results:The results showed that the methanol extracts of Medicago lupulina and Trifolium arvense had strong -glucosidase inhibitory activity with values 5680.01mg ACAEs/g extract and 5254.24mg ACAEs/g extract.Although both plants showed comparable activity for other enzymes, the ethyl acetate extracts exhibited a slightly higher enzyme inhibition profile than that of methanol extract, especially against -glucosidase. Conclusion:The extracts of the plants studied may possess significant therapeutic potential and could be particularly advantageous in the pharmaceutical industry, especially in terms of their inhibition activity on the glucosidase enzyme.

Abstract Phlomis species have traditionally been employed for the treatment of a number of ailments such as influenza, the common cold, asthma, bronchitis, coughs, hemorrhoids, ulcers, and gastrointestinal disorders. The goal of this study is to compare the anti‐diabetic, anti‐glaucoma, and anti‐Alzheimer's properties and antioxidant activities of methanol (MEPT) and aqueous (WEPT) extracts from the aerial part of P. tuberosa L. Tandem mass spectrometry and liquid chromatography (LC–MS/MS) were used to investigate the compounds of MEPT and WEPT. The inhibitory effects of MEPT and WEPT on the enzymes acetylcholinesterase (AChE), carbonic anhydrase II (CA II) and α ‐glycosidase were investigated and molecular docking studies demonstrated binding interactions between the major compounds chlorogenic acid, cyanidin‐3‐ O ‐glycoside, quinic acid, rosmarinic acid with AChE, CA II and α ‐glycosidase enzymes. In addition to that, the total content of phenols and flavonoids was analyzed. Furthermore, the antioxidant activity was evaluated by DPPH, ABTS, DMPD, ferrous ions (Fe 2+ ) chelating, FRAP, CUPRAC, and ferric ions (Fe 3+) reducing methods. This study is the first of its kind to show that MEPT and WEPT possess inhibitory effects on AChE, CA II and α ‐glycosidase, as well as displaying antioxidant properties.

The aerial parts of Helichrysum armenium subsp. armenium were subjected to methanol, ethyl acetate and n-hexane extraction. The extracts were evaluated for their in vitro enzyme inhibitory activity [lipoxygenase (LOX), tyrosinase and elastase], antioxidant activity and toxicity (on fibroblast cells). All the extracts displayed a weak effect on LOX and elastase. The ethyl acetate extract showed the highest inhibition on tyrosinase enzyme with IC50 = 460 µg/mL, while kojic acid had IC50 = 30 µg/mL. The methanol and ethyl acetate extracts compared to the n-hexane extract exhibited stronger DPPH• and ABTS•+ scavenging activity, as well as iron(II)-chelating potential, and were shown to have rich phenolic and flavonoid contents. The methanol extract was toxic to healthy fibroblast cells at a higher concentration with IC50 > 500 µg/mL compared to the other extracts. Phytochemical analysis of the extracts was made quantitatively by LC–MS/MS. The major components identified in the methanol and the ethyl acetate extracts were astragalin, quinic acid, apigenin-7-glycoside, isoquercitrin, chlorogenic acid, naringenin, apigenin and luteolin-7-glucoside. The compounds detected in the n-hexane extract were much fewer. Based on these findings, H. armenium subsp. armenium can be considered to be a natural raw material with the potential to be used in the cosmetic industry.

This study assessed the halophyte species Limonium spathulatum (Desf.) as a possible source of natural ingredients with the capacity to inhibit enzymes related to relevant human health disorders and food browning. Extracts using food-grade solvents such as water and ethanol were prepared by maceration from dried L. spathulatum leaves. They were evaluated for in vitro inhibition activity of enzymes such as acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), α-glucosidase, tyrosinase and lipase, related to Alzheimer's disease, type-2-diabetes mellitus, skin hyperpigmentation, and obesity, respectively. These extracts were also appraised for in vitro acute toxicity on tumoral and non-tumoral cell lines and their chemical composition by high-performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS/MS). The extracts were more effective towards BChE than AChE. The best results were obtained with the hydroethanolic and water extracts, with IC50 values of 0.03 mg/mL and 0.06 mg/mL, respectively. The hydroethanolic extract had the highest capacity to inhibit α-glucosidase (IC50: 0.04 mg/mL), higher than the positive control used (acarbose, IC50 = 3.14 mg/mL). The ethanol extract displayed the best inhibitory activity against tyrosinase (IC50 = 0.34 mg/mL). The tested samples did not inhibit lipase and exhibited low to moderate cytotoxic activity against the tested cell lines. The hydroethanolic extract had a higher diversity of compounds, followed by the ethanol and water samples. Similar molecules were identified in all the extracts and were mainly hydroxybenzoic acids, hydroxycinnamic acids, and flavonoids. Taken together, these results suggest that L. spathulatum should be further explored as a source of bioactive ingredients for the food, cosmetic, and pharmaceutical industries.

Determining the antioxidant abilities and enzyme inhibition profiles of medicinally important plants and their oils is of great importance for a healthy life and the treatment of some common global diseases. Kiwifruit (Actinidia deliciosa) oil was examined and researched using several bioanalytical methods comprehensively for the first time in this research to determine its antioxidant, antiglaucoma, antidiabetic and anti-Alzheimer's capabilities. Additionally, the kiwifruit oil inhibitory effects on acetylcholinesterase (AChE), carbonic anhydrase II (CA II), and α-amylase, which are linked to a number of metabolic illnesses, were established. Furthermore, LC-HRMS analysis was used to assess the phenolic content of kiwifruit oil. It came to light that kiwifruit oil contained 26 different phenolic compounds. According to the LC-HRMS findings, kiwifruit oil is abundant in apigenin (74.24 mg/L oil), epigallocatechin (12.89 mg/L oil), caryophyllene oxide (12.89 mg/L oil), and luteolin (5.49 mg/L oil). In addition, GC-MS and GC-FID studies were used to ascertain the quantity and chemical composition of the essential oils contained in kiwifruit oil. Squalene (53.04%), linoleoyl chloride (20.28%), linoleic acid (2.67%), and palmitic acid (1.54%) were the most abundant compounds in kiwifruit oil. For radical scavenging activities of kiwifruit oil, 1,1-diphenyl-2-picryl-hydrazil (DPPH•) and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS•+) radicals scavenging techniques were examined. These methods effectively demonstrated the potent radical scavenging properties of kiwifruit oil (IC50: 48.55 μg/mL for DPPH•, and IC50: 77.00 μg/mL for ABTS•+ scavenging). Also, for reducing capabilities, iron (Fe3+), copper (Cu2+), and Fe3+-2,4,6-tri(2-pyridyl)-S-triazine (TPTZ) reducing abilities were studied. Moreover, kiwifruit oil showed a considerable inhibition effect towards hCA II (IC50: 505.83 μg/mL), AChE (IC50: 12.80 μg/mL), and α-amylase (IC50: 421.02 μg/mL). The results revealed that the use of kiwifruit oil in a pharmaceutical procedure has very important effects due to its antioxidant, anti-Alzheimer, antidiabetic, and antiglaucoma effects.

Structure-guided design and development of vanillin-triazole conjugates as potential MARK4 inhibitors targeting hepatocellular carcinoma