Antiglycation Effects Research Articles

α-Dicarbonyl compounds trapping ability and antiglycative effect of high-molecular-weight brewer's spent grain melanoidins

Polyphenols participate in the Maillard reaction pathways scavenging α-dicarbonyl compounds (DCs) and contributing to the mitigation of carbonyl burden through dietary exposure/routes. The current study demonstrated the effectiveness of high-molecular-weight brewer's spent grain melanoidins (HMW-BSGM) in reacting with DCs in an in vitro model system. HMW-BSGM (4 mg/mL) quenched more than 95% of glyoxal and methylglyoxal, and more than 80% of 2,3-butanedione after a 7-day incubation at 37 °C. Among tested polyphenols, sinapic acid showed the highest trapping capacity with inhibition rates of 33.1, 49.1 and 49.3% for glyoxal, methylglyoxal and 2,3-butanedione because of hydroxyalkylation reaction as revealed by liquid chromatography high-resolution tandem mass spectrometry experiments. The formation of free fluorescent AGEs was substantially hindered (79.3%) by HMW-BSGM (4 mg/mL). These findings corroborate the hypothesis that the accumulation of polyphenols in melanoidins skeleton can hinder undesired effects and potentially harmful reactions involving α-dicarbonyl compounds.

Polysaccharides from banana (Musa spp.) blossoms: Isolation, identification and anti-glycation effects

Musa spp. (banana) is consumed globally as a healthy fruit and improves the immune system. Banana blossoms are a by-product of banana harvesting rich in active substances such as polysaccharides and phenolic compounds; however, these blossoms are typically discarded as waste. In this report, a polysaccharide, MSBP11, was extracted, purified and identified from banana blossoms. MSBP11 is a neutral homogeneous polysaccharide with a molecular mass of ∼214.43 kDa and composed of arabinose and galactose at a ratio of 0.303:0.697. MSBP11 exhibited potent antioxidant and anti-glycation activities in a dose-dependent manner and can be used as a potential natural antioxidant and inhibitor of advanced glycosylation end products (AGEs). In addition, banana blossoms have been shown to decrease the levels of AGEs in chocolate brownies, which might possibly be developed as functional foods for diabetic patients. This study provides a scientific basis to further research the potential application of banana blossoms in functional foods.

More Than a Diamine Oxidase Inhibitor: L-Aminoguanidine Modulates Polyamine-Related Abiotic Stress Responses of Plants

L-aminoguanidine (AG) is an inhibitor frequently used for investigating plant abiotic stress responses; however, its exact mode of action is not well understood. Many studies used this compound as a specific diamine oxidase inhibitor, whereas other studies used it for reducing nitric oxide (NO) production. Recent studies suggest its antiglycation effect; however, this remains elusive in plants. This review summarises our current knowledge about different targets of AG in plants. Our recommendation is to use AG as a modulator of polyamine-related mechanisms rather than a specific inhibitor. In the future overall investigation is needed to decipher the exact mechanisms of AG. More careful application of AG could give more insight into plant abiotic stress responses.

New Anti-Glycative Lignans from the Defatted Seeds of Sesamum indicum.

Seven known analogs, along with two previously undescribed lignan derivatives sesamlignans A (1) and B (2), were isolated from a water-soluble extract of the defatted sesame seeds (Sesamum indicum L.) by applying the chromatographic separation method. Structures of compounds 1 and 2 were elucidated based on extensive interpretation of 1D, 2D NMR, and HRFABMS spectroscopic data. The absolute configurations were established by analyzing the optical rotation and circular dichroism (CD) spectrum. Inhibitory effects against the formation of advanced glycation end products (AGEs) and peroxynitrite (ONOO-) scavenging assays were performed to evaluate the anti-glycation effects of all isolated compounds. Among the isolated compounds, (1) and (2) showed potent inhibition towards AGEs formation, with IC50 values of 7.5 ± 0.3 and 9.8 ± 0.5 μM, respectively. Furthermore, the new aryltetralin-type lignan 1 exhibited the most potent activity when tested in the in vitro ONOO- scavenging assay.

Abstract 2534: Evaluation of the antiglycation effect and AGEs-protein cross-link breakage ability of cinnamon stem-bark water extract

Abstract 2534: Evaluation of the antiglycation effect and AGEs-protein cross-link breakage ability of cinnamon stem-bark water extract

Vanillic Acid Inhibited the Induced Glycation Using In Vitro and In Vivo Models.

Glycation is implicated in the pathophysiology of many diseases, including diabetes, cancer, neurodegenerative diseases, and aging. Several natural and synthetic compounds were investigated for their antiglycation activity. We evaluated the antiglycation effect of vanillic acid (VA) using in vitro and in vivo experimental models. In vitro, bovine serum albumin (BSA) (50 mg/ml) was incubated with glucose (50 mM) with or without VA at 1.0-100 mM for 1 week at 37°C, and then, excitation/emission fluorescence was measured at 370/440 nm to determine glycation inhibition. The cytoprotective effect of VA was evaluated using RAW 264.7 cells incubated with or without VA at 7.8-500 μM along with 100-400 μM of methylglyoxal for 48 hours, and cell viability was determined using the MTT assay. Aminoguanidine (AMG) was used as a positive control in both in vitro and cell culture experiments. In vivo, 52 streptozotocin-induced diabetic rats were randomly assigned to 4 groups and treated with 0, 1.5, 4.5, or 15 mg/kg VA for four weeks. Serum fructosamine and blood glycosylated hemoglobin (HbA1c) were then measured, and advanced glycation end-products (AGEs) were detected in the kidneys and the skin of deboned tails using an immunohistochemistry assay. VA caused a concentration-dependent effect against BSA glycation (IC50 of 45.53 mM vs. 5.09 mM for AMG). VA enhanced cell viability at all concentrations of VA and methylglyoxal. VA did not affect serum fructosamine or blood HbA1c levels, although it markedly decreased AGEs in the kidney in a dose-dependent manner and decreased AGEs in the skin of deboned tail tissues. VA had significant antiglycation activity at cellular and long-term glycation.

Therapeutic Efficacy of Natural Product 'C-Phycocyanin' in Alleviating Streptozotocin-Induced Diabetes via the Inhibition of Glycation Reaction in Rats.

Diabetes is a long-term metabolic disorder characterized by persistently elevated blood sugar levels. Chronic hyperglycemia enhances glucose-protein interactions, leading to the formation of advanced glycation end products (AGEs), which form irreversible cross-links with a wide variety of macromolecules, and accumulate rapidly in the body tissues. Thus, the objective of this study was to assess the therapeutic properties of C-phycocyanin (C-PC) obtained from Plectonema species against oxidative stress, glycation, and type 2 diabetes mellitus (T2DM) in a streptozotocin (STZ)-induced diabetic Wistar rat. Forty-five days of C-PC administration decreased levels of triglycerides (TGs), blood glucose, glycosylated hemoglobin, (HbA1c), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), liver and kidney function indices, and raised body weight in diabetic rats. C-PC suppressed biochemical glycation markers, as well as serum carboxymethyllysine (CML) and fluorescent AGEs. Additionally, C-PC maintained the redox state by lowering lipid peroxidation and protein-bound carbonyl content (CC), enhancing the activity of high-density lipoprotein cholesterol (HDL-C) and renal antioxidant enzymes, and preserving retinal and renal histopathological characteristics. Thus, we infer that C-PC possesses antidiabetic and antiglycation effects in diabetic rats. C-PC may also act as an antidiabetic and antiglycation agent in vivo that may reduce the risk of secondary diabetic complications.

In Vitro Antioxidant Properties and Methylglyoxal (MGO) Scavenging Effects of Centella asiatica Leaves in Water Extract

Introduction: Advanced glycation end product (AGE) is formed by the non-enzymatic glycation, leading to free radicals’ production and high oxidative stress. Accumulation of AGEs is associated with chronic diseases which are correlated with diabetes mellitus. Centella asiatica is a traditional medicinal herb used in Asia with numerous pharmacological effects such as antioxidant, wound healing, neuroprotective, and gastrointestinal treatment. This research was aim to evaluate the antioxidant, antiglycation, and phytochemical analysis of C. asiatica leaves (L.) water extract. Methods: The antioxidant activity of the plant extract was evaluated through 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (OH) radicals, nitric oxide (NO), and chelating capacity assays. Total phenolic content (TPC) was performed to determine the presence of bioactive compounds, such as saponins, which have been extensively reported in previous studies. Butylated hydroxytoluene (BHT) and ethylenediaminetetraacetic acid (EDTA) were used as positive control in the antioxidant assays, while quercetin was used in the antiglycation assays. Antiglycation effects of C. asiatica L. were evaluated through Bovine serum albumin (BSA)-MGO, BSA-glucose, and MGO scavenging assay. The data was expressed in terms of IC50. Result: The plant extract possessed significantly (p < 0.001) lower IC50 value compared to the BHT in DPPH and NO radical scavenging assay. It also showed stronger antiglycative effect with lower IC50 value compared to the quercetin in BSA-glucose and BSA-MGO assay. The TPC of C. asiatica L. was 3.20 mg/GAE/g at 100µg/ml. Conclusion: The antiglycation effects of C. asiaitca L. include the reduction of Amadori products formation, trapping α-dicarbonyl intermediates, and free radical scavenging. It is suggested that the antiglycation effects of C. asiatica L. maybe attributed by the presence of saponins and other phytochemicals.

Antiglycoxidative properties of amantadine – a systematic review and comprehensive in vitro study

An important drug used in the treatment of Parkinson’s disease is amantadine. We are the first to perform a comprehensive study based on various glycation and oxidation factors, determining the impact of amantadine on protein glycoxidation. Sugars (glucose, fructose, galactose) and aldehydes (glyoxal, methylglyoxal) were used as glycation agents, and chloramine T was used as an oxidant. Glycoxidation biomarkers in albumin treated with amantadine were generally not different from the control group (glycation/oxidation factors), indicating that the drug did not affect oxidation and glycation processes. Molecular docking analysis did not reveal strong binding sites of amantadine on the bovine serum albumin structure. Although amantadine poorly scavenged hydroxyl radical and hydrogen peroxide, it had significantly lower antioxidant and antiglycation effect than all protein oxidation and glycation inhibitors. In some cases, amantadine even demonstrated glycoxidant, proglycation, and prooxidant properties. In summary, amantadine exhibited weak antioxidant properties and a lack of antiglycation activity.

Research Advances on the Damage Mechanism of Skin Glycation and Related Inhibitors.

Our skin is an organ with the largest contact area between the human body and the external environment. Skin aging is affected directly by both endogenous factors and exogenous factors (e.g., UV exposure). Skin saccharification, a non-enzymatic reaction between proteins, e.g., dermal collagen and naturally occurring reducing sugars, is one of the basic root causes of endogenous skin aging. During the reaction, a series of complicated glycation products produced at different reaction stages and pathways are usually collectively referred to as advanced glycation end products (AGEs). AGEs cause cellular dysfunction through the modification of intracellular molecules and accumulate in tissues with aging. AGEs are also associated with a variety of age-related diseases, such as diabetes, cardiovascular disease, renal failure (uremia), and Alzheimer's disease. AGEs accumulate in the skin with age and are amplified through exogenous factors, e.g., ultraviolet radiation, resulting in wrinkles, loss of elasticity, dull yellowing, and other skin problems. This article focuses on the damage mechanism of glucose and its glycation products on the skin by summarizing the biochemical characteristics, compositions, as well as processes of the production and elimination of AGEs. One of the important parts of this article would be to summarize the current AGEs inhibitors to gain insight into the anti-glycation mechanism of the skin and the development of promising natural products with anti-glycation effects.

Effect of crocin on glycated human low-density lipoprotein: A protective and mechanistic approach

Human low-density lipoprotein (LDL) is known to have a role in coronary artery diseases when it undergoes modification due to hyperglycaemic conditions. Plant products like crocin play an essential role in protecting against oxidative stress and in the production of advanced glycation end-products (A.G.E.s). In this study, the anti-glycating effect of crocin was analyzed using various biochemical, spectroscopic, and in silico approaches. Glycation-mediated oxidative stress was confirmed by nitroblue tetrazolium, carbonyl content, and lipid peroxidation assays, and it was efficiently protected by crocin in a concentration-dependent manner. A.N.S. fluorescence, thioflavin T (ThT) assay, and electron microscopy confirmed that the structural changes in LDL during glycation lead to the formation of fibrillar aggregates, which can be minimized by crocin treatment. Moreover, secondary structural perturbations in LDL were observed using circular dichroism (CD) and Fourier transform infrared spectroscopy (FTIR), where crocin was found to prevent the loss of secondary structure in glycated LDL. Spectroscopic studies like U.V. absorbance, fluorescence spectroscopy, CD, FTIR, and fluorescence resonance energy transfer (FRET) provided insights into the interaction mechanism between LDL and crocin. Molecular docking supports these results with a highly negative binding energy of −10.3 kcal/mol, suggesting the formation of a stable ldl-crocin complex. Our study indicates that crocin may be a potent protective agent against coronary artery diseases by limiting the glycation of LDL in people with such disorders.

Antiglycation Effects of Adlay Seed and Its Active Polyphenol Compounds: An In Vitro Study.

This study aimed to evaluate the antiglycation effects of adlay on protein glycation using in vitro glycation assays. Adlay seed was divided into the following four parts: the hull (AH), testa (AT), bran (AB), and polished adlay (PA). A solvent extraction technique and column chromatography were utilized to investigate the active fractions and components of adlay. Based on a BSA-glucose assay, the ethanolic extracts of AT (ATE) and AB (ABE) revealed a greater capacity to inhibit protein glycation. ATE was further consecutively partitioned into four solvent fractions with n-hexane, ethyl acetate (ATE-Ea), 1-butanol (ATE-BuOH), and water. ATE-BuOH and -Ea show marked inhibition of glucose-mediated glycation. Medium–high polarity subfractions eluted from ATE-BuOH below 50% methanol with Diaion HP-20, ATE-BuOH-c to -f, exhibited superior antiglycation activity, with a maximum inhibitory percentage of 88%. Two phenolic compounds, chlorogenic acid and ferulic acid, identified in ATE-BuOH with HPLC, exhibited potent inhibition of the individual stage of protein glycation and its subsequent crosslinking, as evaluated by the BSA-glucose assay, BS-methylglyoxal (MGO) assay, and G.K. peptide-ribose assay. In conclusion, this study demonstrated the antiglycation properties of ATE in vitro that suggest a beneficial effect in targeting hyperglycemia-mediated protein modification.

Beta vulgaris var. cicla methanol extract prevents the formation of advanced glycation end products and protein oxidation against -glucose, and -fructose-induced protein glycation in vitro

Beta vulgaris is an edible plant and herbal medicine traditionally used to treat diabetes and breast cancer and protect against liver damage induced by an HF diet. The aim of this investigation to evaluate the inhibitory activity of B. vulgaris (CH) extract on the formation of advanced glycation end products (AGEs) against fluorescent and Non-fluorescent AGEs, protein oxidation, and protein aggregation in an in vitro model of glucose and fructose protein glycation and Congo red test. Our study provides evidence that CH significantly reduced fluorescent and no fluorescent (Nε-CML) and fructosamine levels in AGEs-BSA systems during four weeks of study. It also avoided protein oxidation, which was shown by the depletion of protein thiol and reduced protein carbonyl, preventing structural loss of protein and AGEs formation by inhibiting protein oxidation and protein glycation. Furthermore, CH extract also inhibited amyloid cross β-structure and increased GPx enzymatic activity. This study demonstrated that CH has a noticeable antiglycation effect in a varied glycation modification of albumin and can retard or forewarn AGEs-related diabetes type 2.

Phytochemical profile, anti-glycation effect, and advanced glycation end-products protein cross-link breaking ability of Sclerocarya birrea stem-bark crude extracts

Introduction: Sclerocarya birrea stem-bark is widely used for the treatment of many medical conditions. Advanced glycation end-products (AGEs) are implicated in the pathogenesis of vascular complications of diabetes mellitus. The study, other than phytochemical composition, evaluated the anti-glycation and AGEs-protein cross-link breaking effects of S. birrea stem-bark extracts. Methods: Different S. birrea extracts and aminoguanidine (used as control) were incubated with bovine serum albumin (BSA) and glucose/fructose at 37oC for 40 days. Amounts of fluorescent AGEs (FAGEs) and immunogenic AGEs formed were determined. Anti-glycation activity percentage of each extract and aminoguanidine was calculated. Their AGEs-protein cross-link breaking abilities were also assessed. Standard techniques were employed for phytochemical screening. Volatile compounds were identified by means of gas chromatography mass spectrometry (GC-MS). Results: S. birrea stem-bark n-hexane extract was statistically more effective than aminoguanidine against the formation of total immunogenic AGEs (P<0.05). For FAGEs, ethyl acetate, methanol, and water extracts exerted significantly higher anti-glycation effects than aminoguanidine (P<0.001). Methanol extract exhibited the highest anti-glycation effect with an average IC50 value of 0.142 mg/mL against FAGEs. All extracts were effective in releasing BSA from the preformed collagen-AGEs-BSA cross-links. GC-MS enabled the identification of many biologically important compounds, including campesterol, stigmasterol, and 1-heptatricontanol. Conclusion: S. birrea stem-bark has a potential for usage in the management of complications in uncontrolled glucose metabolism.

Properties of Cephalopod Skin Ommochromes to Inhibit Free Radicals, and the Maillard Reaction and Retino-Protective Mechanisms in Cellular Models Concerning Oxidative Stress, Angiogenesis, and Inflammation

Ommochromes are pigments of invertebrates that exhibit oxidative stress protection. The aim of this study was to investigate ommochromes extracted from cephalopod’s skin for their ability to inhibit age-related-macular degeneration (AMD)-related factors such as H2O2-induced and iron-dependent oxidative stress (ferroptosis and erastin), accumulation of advanced glycation end-products (AGEs), as well as vascular endothelial growth factor (VEGF), and inflammatory cytokines (interleukin 6 and interleukin 8) secretion. As cell systems, we used primary porcine retinal pigment epithelium (RPE), human retinal pigment epithelium cell line ARPE-19 and uveal melanoma cell line OMM-1. In vitro, ommochromes produced an antiglycation effect by the inhibition of fructosylation reaction. The ommochromes showed protective effects against erastin- induced cell death in ARPE-19. In addition, in long-term stimulation (7 days) ommochromes decreased constitutively secreted VEGF, as well as interleukin 6 and interleukin 8 induced by Poly I:C in primary RPE. No relevant effects were detected in OMM-1 cells. The effects are dependent on the cell system, time of exposition, and concentration. This substance is of interest for further research concerning age-related macular degeneration.

Anti-glycation level of pectic oligosaccharide in orange peel and its stability in accelerated storage temperature

In this study, pectic oligosaccharides of orange peel (OPOs) were isolated and their structure characterized, and then screened according to anti-glycation level compared with aminoguanidine (AG). The results indicated that OPOs mainly included two components, and the main component has more than 50 % inhibition level for the seven glycation products at 60 °C for 40 h. At the accelerated storage temperature, OPOs demonstrated a better anti-glycation level than AG, and this inhibition was concentration-dependent. In addition, the main component in OPOs was separated into 10 fractions by DEAE Sephadex A-25 gel chromatography, the group of 5–7 monosaccharide polymerization showed the best anti-glycation effect, average anti-glycation capability on six products was over 70 % in the 40th hour. The anti-glycation level of this group was closely related to its high content of GalA and molar ratio of GalA: Rha, and positively correlated with time at moderate temperature.

Nutraceutical potential of navel orange peel in diabetes management: The chemical profile, antioxidant, α-glucosidase inhibitory and antiglycation effects of its flavonoids

Oxidative damage, chronic hyperglycemia, and the formation of advanced glycation end products (AGEs) are important risk factors associated closely with diabetes and its complications. In the present study, the antidiabetic potential of navel orange peel was explored for the first time by evaluating the antioxidant, α-glucosidase inhibition, and antiglycation activities of its extracts and individual flavonoids. In total, seventeen flavonoids were identified from the peel extract, of which 5-hydroxy-6,7,3′,4′-tetramethoxyflavone, didymin and sinensetin were the most potent antioxidant, α-glucosidase inhibitor, and glycation inhibitor, respectively. In addition, the structure-activity relationship (SAR) studies of these flavonoids were performed in detail. Moreover, the most potent antidiabetic extract of navel orange peel was quantitatively analyzed by HPLC. The present results support the potent application of orange peel as a nutraceutical resource for diabetes management, and might be beneficial to the orange juice industry by providing useful information on the valorization of orange peel.

Development of a Herbal Tea with Potential Antiglycation Effects using <em>Phyllanthus emblica</em> (Indian Gooseberry), <em>Zingiber officinale</em> (Ginger), and <em>Coriander sativum</em> (Coriander)

The study was conducted to investigate the capability of developing a herbal tea using Phyllanthus emblica (PE) fruit, Zingiber officinale (ZO) rhizome, and Coriander sativum (CS) seeds and to assess its anti-glycation effects. Phyllanthus emblica, ZO, and CS are common and individually used materials in traditional medicine in Sri Lanka. No evidence is found in using combinations of these plant materials being used in commercial tea / herbal infusion production in Sri Lanka. The herbal tea was formulated with powdered form of PE fruits, ZO rhizome and CS seeds in three different formulations; PE-50% + ZO-25% + CS-25%; PE-25% + ZO-50% + CS-25%; and PE-25% + ZO-25% + CS-50%. The three formulated teas were subjected to a five-point hedonic scale sensory evaluation. The formula with PE fruits 50%, ZO rhizome 25%, and CS seeds 25% was selected for further analysis. The phytochemicals of the selected formulation were analyzed, and the results bared the presence of glycosides, alkaloids, terpenoids, saponins, and tannins, which ensures a high antioxidant quality of the formulated tea. The tested parameter for the shelf-life determination study concluded that the product is shelf-stable for 14 days under 0 °C temperature. An in vitro assay using Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis was used to test the antiglycation effects. Results showed that the herbal tea was able to inhibit glycation-induced protein cross-linking over prolonged incubation periods with strong glycating conditions. Therefore, this product could also have a potential to be used as a home remedy to prevent diabetic complications.

Combination of LC-Q-TOF-MS/MS, network pharmacology, and nanoemulsion approaches identifies active compounds of two Artemisia species responsible for tackling early diabetes-related metabolic complications in the liver.

The chronicity of advanced glycation end-products (AGEs) imparts various damages resulting in metabolic dysfunction and diseases involving inflammation and oxidative stress. The use of plant extracts is of high interest in complementary medicine. Yet, extracts are multicomponent mixtures, and difficult to pinpoint their exact mechanism. We hypothesise that network pharmacology and bioinformatics can help experimental findings depict the exact active components and mechanism of action by which they induce their effects. Additionally, the toxicity and variability can be lowered and standardised with proper encapsulation methods. Here, we propose the formulation of phytoniosomes encapsulating two Artemisia species (Artemisia dracunculus and Artemisia absinthium) to mitigate AGEs and their induced cell redox dysregulation in the liver. Extracts from different solvents were identified via liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS/MS). Phytoniosomes were explored for their anti-glycating effect and modulation of AGE-induced damages in THLE-2 liver cells. Network pharmacology tools were used to identify possible targets and signalling pathways implicated. Data demonstrated that A. absinthium phytoniosomes had a significant anti-AGE effect comparable to reference molecules and higher than A. dracunculus. They were able to restore cell dysfunction through the restoration of tumour necrosis alpha (TNF-α), interleukin 6 (IL-6), nitric oxide, and total antioxidant capacity. Phytoniosomes were able to protect cells from apoptosis by decreasing caspase 3 activity. Network pharmacology and bioinformatic analysis confirmed the induction of the effect via Akt-PI3K-MAPK and AGE-RAGE signalling pathways through quercetin and luteolin actions. The current report highlights the potential of Artemisia phytoniosomes as strong contenders in AGE-related disease therapy.

Oleuropein-Enriched Extract From Olive Mill Leaves by Homogenizer-Assisted Extraction and Its Antioxidant and Antiglycating Activities.

Olive oil consumption has increased in the last two decades and consequently, its wastes have increased, which generates a tremendous environmental impact. Among the by-products are the olive mill leaves, which are easier and inexpensive to treat than other olive by-products. However, little research has been done on their chemical composition and potential bioactivity. Hence, in this study, olive mill leaves were used to obtain Oleuropein-Enriched Extracts (OLEU-EE) using Conventional Extraction, Ultrasound-Assisted Extraction, and Homogenization-Assisted Extraction. These three techniques were evaluated using a Factorial Design to determine the parameters to obtain an OLEU-EE with high contents of Total Phenolic Compounds (TPC), Antioxidant Activity (AA), and Oleuropein concentration (OLEU). From the results, the Homogenizer-Assisted Extraction (HAE) technique was selected at 18,000 rpm, solid:liquid ratio 1:10, and 30 s of homogenization with 70% ethanol, due to its high TPC (5,196 mg GA/100 g), AA (57,867 μmol of TE/100 g), and OLEU (4,345 mg of OLEU/100 g). In addition, the antiglycating effect of OLEU-EE on the levels of (1) fluorescent Advanced Glycation End Products (AGEs) were IC50 of 0.1899 and 0.1697 mg/mL for 1λEXC 325/λEM 440 and 2λEXC 389/λEM 443, respectively; (2) protein oxidative damage markers such as dityrosine (DiTyr), N-formylkynurenine (N-formyl Kyn), and kynurenine (Kyn) were IC50 of 0.1852, 0.2044, and 0.1720 mg/mL, respectively. In conclusion, OLEU-EE from olive mill leaves has different capacities to inhibit AGEs evidenced by the IC50 of fluorescent AGEs and protein oxidation products, together with the scavenging free radical evidenced by the concentration of Trolox Equivalent. Therefore, OLEU-EE could be potential functional ingredients that prevent oxidative damage caused by free radicals and AGEs accumulation.