Alpha-Amylase Inhibition by Myricetin Nanoparticles: An in Silico, In Vitro, and In Vivo Evaluation toward Natural Antidiabetic Therapy

Alpha-Amylase Inhibition by Myricetin Nanoparticles: An in Silico, In Vitro, and In Vivo Evaluation toward Natural Antidiabetic Therapy | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Alpha-Amylase Inhibition by Myricetin Nanoparticles: An in Silico, In Vitro, and In Vivo Evaluation toward Natural Antidiabetic Therapy Karishma Waghmare, Guno Sindhu Chakraborthy This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8680908/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Purpose Diabetes mellitus is a worldwide health challenge, concern, and effective and affordably priced therapies are pivotal. Myricetin, a naturally occurring flavanol, has demonstrated promising antidiabetic potential. The current research aims to investigate the binding affinity and molecular interactions of myricetin with alpha-amylase and 1OSE protein, and to explore its potential as a therapeutic agent for diabetes management. Methods The present investigation, allied with molecular docking of myricetin against diabetes related targets, was investigated through Auto dock Vina, PyRx, and Schrodinger Glide. The target proteins were prepared in PDBQT format. Myricetin was docked to explore its molecular mechanism. The binding affinity and orientation of myricetin in the active site of alpha-amylase and 1OSE protein were evaluated. The in vivo antidiabetic potential of myricetin nanoparticles was assessed in Streptozotocin-induced diabetic Wistar rats. The experimental protocol was approved by the Institutional Animal Ethics Committee (IAEC) of Invitox R and D institute registration no.2273/PO/RERC/S/23 CCSEA, Report no. IRDI/IVAS/18/2024-25 registered under CPCSEA. The animals were randomly assigned to five groups (n = 6). Group I served as the normal control, while Group II received intraperitoneal Streptozotocin and was maintained as the untreated diabetic control. Groups III and IV were administered the test formulation (F1) orally at the selected doses, whereas Group V received the standard antidiabetic drug glibenclamide (5 mg/kg). The F1-treated groups exhibited a significant decrease in fasting blood levels of glucose compared with the diabetic control group, confirming in vivo antidiabetic efficacy of myricetin nanoparticles. Results Myricetin exhibited a strong and stable binding affinity toward alpha-amylase and 1OSE protein, with a binding energy of -8.8 to -7.6 kcal/mol. Molecular docking results revealed multiple hydrogen bonds and hydrophobic contacts between myricetin and critical residues within the catalytic pocket. Conclusion Myricetin exhibits promise as a therapeutic propensity for diabetes mitigation owing to its strong binding affinity and inhibitory activity against alpha-amylase. Alpha-amylase diabetes in silico in vitro molecular docking myricetin Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Introduction Diabetes mellitus, an endocrine disorder characterized by metabolic dysregulation and high blood sugar levels, presents symptoms such as the excessive urination (polyuria); smell of acetone on the patient's breath; rapid, deep breathing (Kussmaul breathing); excessive thirst (polydipsia); excessive hunger (polyphagia); nausea; vomiting; abdominal pain; blurred vision; weight loss; and various altered states of consciousness or arousal (including hostility, mania, confusion, and lethargy). This condition poses perils such as cardiovascular diseases, neuropathy, nephropathy, and retinopathy [ 1 , 2 ]. Type I, Type II, gestational, maturity onset at a young age, and malnutrition-related are the types of diabetes[ 3 ].Several factors attribute to the development of diabetes, viz, impaired responsiveness to insulin by body’s cells, reduced glucose uptake, impaired insulin secretion owing to dysfunction, destruction of pancreatic beta cells, scarcity in insulin production, glucose imbalance, other attributing factors like genetics, obesity, scarcity of physical activity, unhealthy diet, and age[ 4 , 5 ]. Regular exercise, a healthy diet, weight loss, and stress management avert diabetes[ 6 , 7 ]. If left untreated, it leads to perils, viz., heart ailments, stroke, peripheral artery disease, kidney damage (even failure may occur), nerve damage leading to numbness, tingling, pain, eye damage, encompassing blindness, and impediments in hearing. Complications of diabetes like cancer, end stage kidney disease ,peripheral artery disease found globally. The high rate of mortality is due to macrovasular and microvascular complications which affects quality of life [ 8 , 9 ]. The increased population that is at risk of developing diabetes needs affordable and effective mitigation strategies [ 10 , 11 ].Despite the fact that synthetic drugs continue to play a pivotal role in the mitigation of diabetes, compounds that originate from plants have allured attention as a new and promising therapeutic agent owing to their low toxicity and side effects [ 12 , 13 ].Flavonoids are polyphenolic compounds that are found in abundance in fruits, vegetables, and beverages like tea and wine [ 14 , 15 ] .They are classified in various subgroups, which are flavonols, flavones, flavanones, and isoflavones[ 16 , 17 ]. Myricetin is a flavonol,[ 18 ] which has largely been studied with respect to its numerous biological actions. Myricetin is an antioxidant,[ 19 ] anti-inflammatory,[ 20 , 21 ] antimicrobial,[ 22 , 23 ] and anticancer drug[ 24 ] and thus a very strong pharmaceutical substance in medications for metabolic and degenerative disease, which include diabetes. Myricetin, a flavonoid found in various plants, has been extensively studied for its antioxidant, neuroprotective,[ 25 , 26 ] and pharmacological properties in vitro. Myricetin is a possible therapy option for neurological ailments based on research revealing it can suppress oxidative stress, boost natural antioxidant enzymes, and neutralize free radicals. Flavonoid compounds, including myricetin, generally possess low bioavailability, which may limit their clinical usefulness despite their established high biological activity. Glycosylation, metabolism, and absorption are factors that affect systemic availability [ 27 ]. However, evidence exists to demonstrate that dietary flavonoid is safe in dietary quantities[ 28 ]. Antioxidant and anti-inflammatory activities are significant in averting dysfunction of β-cell and insulin resistance, rather than causing them. Myricetin is common flavonoid available in plants. It is an attenuator of oxidative stress that removes free radicals and amplifies endogenous antioxidant enzymes. Inhibition of α-Glucosidase and α-Amylase Myricetin and other flavonoids prevent the rise of postprandial glucose levels by inhibiting carbohydrate-searching enzymes. This process can be likened to artificial pills such as acarbose. According to the Insulin Mimetic and Secretagogue Action Studies, myricetin boosts glucose intake in an insulin-like way that involves mimicking and hierarchical insulin regulation. It also activates also insulin secretion of pancreatic β-Cell. The neuroprotective potency of myricetin in β-cells of the pancreas has been established through its antioxidative activity which decreases apoptosis and maintains functionality. A few animal experiments elucidate the antidiabetic effect of myricetin. Preclinical studies has shown antihypertensive,antidiabetic ,anticancer and antioxidant activities. [29 ]. Treatments with myricetin enhanced glycemic control and improved the lipid profiles and sensitivity of insulin in diabetic rat. There is some human clinical evidence that supports possible use of polyphenol-rich diets in glycemic control albeit limited. Other Flavonoids Quercetin and the structural analogs luteolin and kaempferol have been able to show antidiabetic effect through similar means, i.e., by enzyme inhibition and anti-inflammatory action. Poor solubility, potency, low bioavailability and the failure to investigate in large scale clinical trials are some of the challenges encountered that may be solved in near future. To address these limitations, nanoparticle delivery systems,[ 30 , 31 ] and glycoside derivatives are under investigation. AutoDock Vina,[ 32 ] PyRx,[ 33 ] and Schrodinger Glide[ 34 ] are employed to run the molecular docking which simulates the small molecule (ligand)-target protein interaction to predict the binding affinity and orientation in the active site. In general, these tools treat the protein receptor as a rigid body, but semi-flexible docking is sometimes permitted, whereas the ligand is assumed to be fully flexible thereby permitting investigation of all rotatable bonds and torsional conformations. The generated poses are scored with a semi-empirical scoring function which estimates binding affinity which is usually reported in kcal/mol, the lower the value of binding energy indicates the higher binding affinity and preferred ligand-receptor interactions, such as hydrogen bonding, van der Waals force, pi pi-stacking and electrostatic interaction, which are usually visualized with a molecular visualization package including Discovery Studio, PyMOL or Chimera. The active site is bounded by a grid box to cut the search space and enhance the docking efficiency. This grid restricts the docking algorithm to a pertinent region on the receptor that increases specificity. The present investigation focuses on assessing the antidiabetic potential of myricetin utilizing both computer-based simulations ( in silico ) and laboratory experiments (in vitro ). Materials and Methods Materials: In silico tools: Various molecular modeling open-source tools such as Discovery Studio, PyMOL or Chimera, Auto dock, [ 35 , 36–38 ] PyRx, [ 33 ] and Schrodinger Glide [39–43 ] etc. were employed in present investigation. Assay chemicals: Myricetin (≥98% purity) was purchased from Yucca Enterprises Mumbai, Porcine pancreatic α-amylase, and Phosphate buffer pH 6.8 were procured from Loba Chemie; Soluble starch from Loba Chemie; All solvents and chemicals were all analytical grades. Deionized water was employed throughout the study. Chemicals: Copper chloride was purchased from Loba Chemie. Preparation of myricetin nanoparticles 0.1 M Copper sulphate dissolved in 100 ml distilled water; stirred the solution to obtain a clear, blue-coloured copper salt solution. Myricetin was slowly added to copper sulphate solution with constant stirring. The mixture was heated at 60–80 °C for 30–60 minutes. Colour change typically from blue to dark brown, was noted indicating nanoparticle formation. Reaction mixture was made to stand for a few hours for complete reduction. Mixture was centrifuged at 8000–10000 rpm for about 15–20 minutes. The purified nanoparticles dried in a hot air oven at 50-60 °C. Characterization of myricetin nanoparticles Physiochemical studies Percent yield of the synthesized derivative, melting point, pH (assess compatibility with biological system), solubility and dispersibility (assess aqueous solubility along with dispersion behaviour) were recorded [44,45]. FTIR Samples were analysed by FTIR spectropho­tometer (Alpha T Bruker) in the standard region 4000–400 cm 1 by potassium bromide method to confirm functional groups, chemical interaction between adjuvants, and nanoparticle components. FTIR spectra were recorded for myricetin nanoparticles. Scans were interpreted for the retention of principle peaks, shifting and new peaks appearance. Figure 1 illustrates FTIR spectra [46,47]. Particle size SEM of was assessed with the help of scanning electron microscope (VEG A3 TESCAN), at 15 keV accelerating voltage. 2 mg samples were coated in vacuum employing thin gold layer before assessment [ 47,48] SEM photographs of myricetin nanoparticles is illustrated in Figure 4. Zeta potential assessment Zeta potential assesses surface charge. Zeta potential was recorded for CMGG and nanoparticles employing a zeta potential analyser (Zetasizer Horiba Scientific, Japan). Figure 3 illustrates zeta potential analysis [48]. In silico molecular docking assessment [ 49 ]. Ligand preparation 3D structure of myricetin was retrieved from PubChem database (CID: 5281672) in SDF format and converted to PDB format employing Open Babel. Ligand was energy-minimized and converted to PDBQT format employing AutoDock Tools, assigning Gasteiger charges and adding rotatable bonds . Protein preparation Crystal structures of target proteins-human pancreatic α-amylase (PDB ID: 1HNY) and α-amylase-acarbose complex (PDB ID: 1OSE) were downloaded from the RCSB Protein Data Bank. Lligands, water molecules, and ions were removed. Polar hydrogens were added, and proteins were converted to PDBQT format. Interaction analysis Ligand-protein interactions were visualized employing Discovery Studio Visualizer 2020. Hydrogen bonding, hydrophobic interactions, and π–π stacking were assessed to identify key residues in ligand binding within the catalytic site. In vitro α -amylase inhibition assay α-amylase inhibition potential of myricetin and its nanoparticles was determined by Dinitro salicylic acid (DNS) method [ 50 ] . Briefly, 500 µL of test sample was allowed to react with 0.1 M 500 µL of 0.5 % α-amylase solution (1 U/mL) in Phosphate buffer solution (PBS) pH 6.9 and incubated at 25 °C for 10 min. Subsequently, 500 µL of 1% starch solution in 0.1 M PBS 6.8 was added and incubated at 25 °C for another 10 min. same procedure was redetected for control where 500 µL of the enzyme was replaced by buffer. Reaction was terminated by adding 1000 µL of DNS reagent, followed by boiling at 90 °C for 5 min. Standard acarbose (α-amylase enzyme inhibitor) was employed as standard drug. After cooling to room temperature, absorbance was measured at 540 nm employing UV spectrophotometer (Lab India 3000). % inhibition of α-amylase enzyme was computed using formula as follows. Streptozotocin induced antidiabetic activity As shown in the table no. 1 ,Wistar rats were divided into 5 groups, with 6 rats in each. The second group was Streptozotocin-induced diabetic injection intra peritoneal and did not undergo any treatment. The diabetic of the second group, used as reference, and that of the third to fifth group was treated by an administered orally test item (F1) and Glibenclamide (5 mg/kg), respectively, to elevate their blood glucose levels. Blood samples were collected from the retro orbital of the overnight (12-15h) fasted rats and blood glucose level was determined on 0th, 7th, 14th and 21st day along with body weight and body temperature. If the blood glucose levels of rats> 200 mg/dL then the rats are considered to have hyperglycaemia. In all treated groups, the glucose level was measured before and after the Streptozotocin-induced diabetic injection using a digital glucometer. At specified time intervals (On day 0, 7, 14, 21 after treatment), blood was collected from retro orbital sinuses. Blood glucose levels were determined using a digital glucometer. Result and discussion Characterization of myricetin nanoparticles The yield of the synthesized myricetin nanoparticles was approximately in the 75-80%, melting point was 220 ºC, pH was 6.8. Myricetin nanoparticles swelled, formed clear dispersion in water with no signs of precipitation or sedimentation after 24 hours suggesting high solubility. Nanoparticles remained uniformly suspended for long duration with negligible sedimentation thus it can be concluded that nanoparticles exhibited good dispersibility. Thus, it was concluded that synthesized myricetin nanoparticles exhibited desired physical and chemical features. FTIR Samples were analysed by FTIR spectropho­tometer (Alpha T Bruker) in the standard region 4000–400 cm 1 to confirm functional groups, chemical interaction between myricetin and copper chloride, and nanoparticle components. Figure 1and 2 illustrates FTIR spectra of myricetin and nanoparticles. Hydroxyl and carbonyl groups (–OH and C=O) present in myricetin and active in binding to metal ions, strong band at 1645 cm⁻¹ confirms carbonyl group. Metal–Oxygen Bond Formation, Peaks at <600 cm⁻¹ (e.g., 518, 472, 408 cm⁻¹) represent Cu–O or Cu–OH bonds, confirms successful loading or formation of copper nanoparticles with plant-derived compounds. Thus, FTIR spectrum confirms the presence of myricetin functional groups (phenolic –OH, carbonyl, etc.) Their involvement in the reduction and stabilization of copper nanoparticles. Formation of Cu–O bonds confirm successful synthesis of myricetin copper nanoparticles through green synthesis routes. Shift in C=O stretch (~1656 → 1645 cm⁻¹) shows coordination of Cu²⁺ with carbonyl oxygen in myricetin. Reduction/shift of OH Stretch (~3291 & 2920 cm⁻¹) indicates hydrogen bonding changes or involvement in metal complexation. Appearance of Cu-O peaks (< 600 cm⁻¹) at 677, 518, 472, 408 cm⁻¹ are absent in pure myricetin, present only in copper nanoparticles spectrum. Thus, it can be concluded from FTIR that there is successful metal-ligand bond formation. Particle size Particle size assessment was employed to determine average size, particle size distribution, homogeneity of nanoparticles and was analysed employing a particle size analyser (Figure 3). The main peak is between 100 and ~300 nm, with most particles clustering around 183–210 nm. Distribution type was single peak and moderate spread. Polydispersity index was 0.58 indicates a broad size distribution, likely due to aggregation. TEM images of myricetin nanoparticle demonstrate spherical shape and are dense nature with clear boundaries with no aggregation. Nanoparticle demonstrates 30-40 nm in diameter; thus, it was concluded that these nanoparticles were in nanometre range. SEM of myricetin nanoparticle also demonstrate its spherical nature. Figure 4a and 3b illustrates TEM and SEM of myricetin nanoparticle respectively. Zeta potential assessment Zeta potential assesses surface charge. Zeta potential was recorded for myricetin nanoparticles employing a zeta potential analyser and was found to be -35 mV hold a moderately negative surface charge and nanoparticles are stable as zeta potential value is closer to ±30 mV. Figure 5 illustrates zeta potential analysis. 3.1 Molecular docking Molecular docking analysis has been performed against two critical diabetes-related proteins α-amylase and 1OSE employing AutoDock, PyRx, and Schrodinger Glide. The least binding affinity represents the strongest binding affinity. Binding affinity (kcal/mol) indicates strength of ligand binding; more negative values suggest stronger binding. Root Mean Square Deviation, upper bound (RMSD/ub) measures deviation from the reference pose. RMSD/lb (lower bound) like ub, typically employed to validate docking accuracy. Lower RMSD values (<2 Å) indicate more reliable binding poses. Figure 6a demonstrates smd-Myricetin 2D and 6b smd Myricetin 3D structures respectively. Figure 7a demonstrates 1OSE-Myricetin 2D and 7b Myricetin 3D structures respectively. Myricetin binds effectively to 1OSE with multiple stable conformations; docking is robust, with several poses under 2 Å RMSD supporting pose stability and reproducibility. Molecular docking results of myricetin against 1OSE protein revealed a highly favourable interaction profile. Top-ranked pose exhibited a binding affinity of -8.8 kcal/mol, suggesting a strong and energetically stable interaction between ligand and protein’s active site. This value reflects a high probability of effective binding under physiological conditions, making myricetin a promising candidate for modulating 1OSE-related activity. Root mean square deviation (RMSD) of this top pose was 0 Å, confirming that it is reference conformation-the most accurate and least deviated pose generated during the docking simulation. RMSD of 0 implies that this pose was used as a baseline for comparing other generated conformations and likely represents the most native-like fit within the binding pocket. Several additional poses demonstrated RMSD values less than 2 Å (e.g., 2.812 Å, 1.690 Å, 1.275 Å), which are within accepted range for reliable docking predictions. Poses with RMSD < 2 Å are generally considered to maintain a similar binding orientation as the reference pose, indicating high reproducibility and consistency of ligand binding across docking iterations. These conformations further validate the robustness of docking protocol and suggest that Myricetin can adopt multiple stable binding geometries in 1OSE active site. In contrast, several poses exhibited significantly higher RMSD values (e.g., 17.073 Å, 17.815 Å, 8.99 Å), suggesting these conformations likely represent alternative binding modes or non-optimal orientations within or near binding cavity. myricetin, a naturally occurring flavonol, exhibits a strong and stable binding affinity toward α-amylase, a key enzyme involved in carbohydrate digestion and postprandial hyperglycaemia. Molecular docking results revealed a highly favourable binding energy, indicating that myricetin can effectively occupy the active site of α-amylase and potentially inhibit its enzymatic activity. Detailed interaction analysis showed that myricetin forms multiple hydrogen bonds and hydrophobic contacts with critical residues within the catalytic pocket, suggesting competitive inhibition as a likely mechanism of action. In vitro α -amylase inhibition assay The digestive enzyme α-amylase is responsible for hydrolysing dietary starch (maltose), which breaks down into glucose prior to absorption. Inhibition of α-amylase can lead to reduction in post prandial hyperglycaemia in diabetic condition. Given sample demonstrated good inhibition of α-amylase enzyme when compared to acarbose standard. Streptozotocin induced antidiabetic activity The results were expressed as mean SD (n=6), ns p>0.05, non- significant; p <0.01, When compared with positive control group. Based on the provide data on glucose levels over the course of the experiment, here is a conclusion drawn regarding the anti-diabetic activity. Table 1 Boold glucose level in diabetic Wistar rat Groups Glucose level on day 0 (mg/dl) Glucose level on day 7 (mg/dl) Glucose level on day 14 (mg/dl) Glucose level on day 21(mg/dl) Normal group 85.98±4.23 88.45±2.15 86.36±3.69 85.25±4.28 Diabetic control 230.97±1.36 220.24±4.28 275.28±5.89 270.18±7.28 Standard 225.3±4.11 258.57±2.03 160.51±0.79 110.68±0.98 Nanoparticles myricetin (100 mg/kg) 227.58±1.36 270.18±4.05 150.18±7.25 95.35±5.20 Blood glucose lowering study (hypoglycemic study) Myricetin nanoparticles showed a significant and consistent reduction in blood glucose levels over 21 days (figure 8) After an initial rise by day 7 due to induced hyperglycaemia, treatment led to a marked decrease by day 14 and near-normal levels by day 21. This improved effect is attributed to enhanced bioavailability and targeted delivery through the nanoparticle formulation. Compared to the simple extract and Glibenclamide, Myricetin nanoparticles demonstrated superior anti-diabetic activity, likely due to their antioxidant and insulin-sensitizing properties. These findings support the potential of Myricetin nanoparticles as an effective anti-diabetic agent. Figure 9 illustrates blood glucose level in diabetic Wistar rat. Conclusion The current investigation infers that myricetin nanoparticles exhibited significant α-amylase inhibitory potential, emphasizing their propensity as a natural antidiabetic entity. Molecular docking assessment revealed strong binding affinity of myricetin to the active site of α-amylase, assisted by favourable interaction energies and key hydrogen bond formations. In vitro α-amylase inhibition assay further supported outcome of the current investigation, with myricetin nanoparticles exhibiting ameliorated inhibition. The blend in silico and in vitro approach highlights the therapeutic promise of myricetin-loaded nanoparticles in mitigating hyperglycemia, presenting a cost-effective and substitute to conventional therapies. Declarations Conflict of Interest: The authors declare no conflict of interest Funding: The authors did not receive support from any organization for the submitted work. Author Contribution Karishma Ramchandra Waghmare: Conceptualization, formal analysis, experimental, methodology, data curation and writing original draft; Guno Sindhu Chakraborthy: Resources, and literature survey; supervision, review and editing. Acknowledgement The authors are grateful to Yucca Enterprises, Mumbai, India, for providing Myricetin used as a marker in this study. The authors sincerely thank ICON LABS, ICON HOUSE, India, and Yashwantrao Chavan Institute of Science, Satara, for their valuable support in carrying out the characterization studies. The authors also extend their gratitude to Invitox R and D Institute, Pune, India, for conducting the in vivo activity. The generous laboratory facilities provided by the Principal, Shri DD Vispute college of pharmacy and Research center New Panvel, India, gratefully acknowledged.Data Availability All data generated or analysed during this study are included in this published article.DeclarationsConflict of Interest : The authors declare no competing interests. References Antar SA, Ashour NA, Sharaky M, et al. Diabetes mellitus: Classification, mediators, and complications; A gate to identify potential targets for the development of new effective treatments. Biomedicine & Pharmacotherapy. 2023;168:115734. doi:10.1016/j.biopha.2023.115734 RK Dey. Diabetes Mellitus: A Comprehensive Review of Pathophysiology, Management, and Emerging Therapeutic Approaches. Journal of Diabetes Medication & Care. 2023;6(4). Ojo OA, Ibrahim HS, Rotimi DE, Ogunlakin AD, Ojo AB. 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Protein Journal. 2013;32(5):356-364. doi:10.1007/s10930-013-9492-3 Shanmuga Priya VG, Swaminathan P, Muddapur UM, Fandilolu PM, Parulekar RS, Sonawane KD. Peptide Similarity Search Based and Virtual Screening Based Strategies to Identify Small Molecules to Inhibit CarD–RNAP Interaction in M. tuberculosis. Int J Pept Res Ther. 2019;25(2):697-709. doi:10.1007/s10989-018-9716-7 Childers MC, Daggett V. Validating Molecular Dynamics Simulations against Experimental Observables in Light of Underlying Conformational Ensembles. Journal of Physical Chemistry B. 2018;122(26):6673-6689. doi:10.1021/acs.jpcb.8b02144 Barage SH, Jalkute CB, Dhanavade MJ, Sonawane KD. Simulated interactions between endothelin converting enzyme and aβ peptide: Insights into subsite recognition and cleavage mechanism. Int J Pept Res Ther. 2014;20(4):409-420. doi:10.1007/s10989-014-9403-2 Barale SS, Parulekar RS, Fandilolu PM, Dhanavade MJ, Sonawane KD. Molecular Insights into Destabilization of Alzheimer’s Aβ Protofibril by Arginine Containing Short Peptides: A Molecular Modeling Approach. ACS Omega. 2019;4(1):892-903. doi:10.1021/acsomega.8b02672 Dhanavade MJ, Sonawane KD. Insights into the molecular interactions between aminopeptidase and amyloid beta peptide using molecular modeling techniques. Amino Acids. 2014;46(8):1853-1866. doi:10.1007/s00726-014-1740-0 Sonawane KD, Dhanavade MJ. Computational approaches to understand cleavage mechanism of amyloid beta (Aβ) peptide. In: Neuromethods. Vol 132. Humana Press Inc.; 2018:263-282. doi:10.1007/978-1-4939-7404-7_11 Chi XQ, Hou B, Yang L, et al. Design, synthesis and cholinesterase inhibitory activity of α-mangostin derivatives. Nat Prod Res. 2020;34(10):1380-1388. doi:10.1080/14786419.2018.1510925 Gupta AP, Verma DK. Synthesis and characterization of carboxymethyl guar gum nanoparticles stabilized polyaniline/carboxymethyl guar gum nanocomposites. J Nanostructure Chem. 2015;5(4):405-412. doi:10.1007/s40097-015-0172-z Dodi G, Pala A, Barbu E, et al. Carboxymethyl Guar Gum Nanoparticles for Drug Delivery Applications: Preparation and Preliminary in-Vitro Investigations. Sreelatha K, AnandaKumar CS, Saraswathi M, Anusha P, Rose NM, Bhargava D. A Comprehensive Review of Nanoparticle Characterization Techniques. International Journal of Research and Review. 2025;12(1):194-200. doi:10.52403/ijrr.20250124 Eid MM. Characterization of Nanoparticles by FTIR and FTIR-Microscopy. In: Handbook /of C onsumer Nanoproducts. Springer Singapore; 2021:1-30. doi:10.1007/978-981-15-6453-6_89-1 Rebecca M, Al Amri Z, Mfon RE. Synthesis, Characterisation and Zeta Potential of Silver Nanoparticles.; 2023. https://www.researchgate.net/publication/368476484 Rathod Shubhangi H. Bhowate. Dinesh R. Chaple DrAJ asnani, PI, AVLVSB. Molecular Docking: A Powerful Tool In Modern Drug Discovery And Its Approaches . Ibrahim MM, Zaki ER, Rady MR. Alpha-amylase inhibitory activity and in silico studies of in vitro sweet basil plantlets treated with chitosan and ZnO NPs. In Vitro Cellular & Developmental Biology - Plant. 2024;60(2):147-160. doi:10.1007/s11627-023-10401-0 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8680908","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":594639823,"identity":"954381e2-de25-4840-b0cc-72fe09cf04a4","order_by":0,"name":"Karishma Waghmare","email":"data:image/png;base64,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","orcid":"","institution":"Parul University","correspondingAuthor":true,"prefix":"","firstName":"Karishma","middleName":"","lastName":"Waghmare","suffix":""},{"id":594639825,"identity":"1b6b89f8-5fd5-48e7-bf09-acfcdc6b7320","order_by":1,"name":"Guno Sindhu Chakraborthy","email":"","orcid":"","institution":"Parul University","correspondingAuthor":false,"prefix":"","firstName":"Guno","middleName":"Sindhu","lastName":"Chakraborthy","suffix":""}],"badges":[],"createdAt":"2026-01-23 15:55:56","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8680908/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8680908/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":104400466,"identity":"3f45b0a4-f365-40d7-a59a-63adb7afb574","added_by":"auto","created_at":"2026-03-11 12:10:03","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":360790,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectra of Myricetin\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8680908/v1/fbe57ba9eca53c1f8ca0f092.png"},{"id":103772277,"identity":"52841bf3-4990-4395-a85d-f933549104b5","added_by":"auto","created_at":"2026-03-02 17:32:29","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":292347,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectra of myricetin nanoparticles\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8680908/v1/3bd7a9ec665c95d63494c72a.png"},{"id":103772272,"identity":"934f7de0-1e67-426b-8d2e-0631050da64d","added_by":"auto","created_at":"2026-03-02 17:32:29","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":77726,"visible":true,"origin":"","legend":"\u003cp\u003eParticle size analysis of myricetin nanoparticle by HORIBA particle size analyser\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-8680908/v1/79f1ae2d120b50edb5098751.png"},{"id":104400343,"identity":"24217997-69c2-412a-b84c-5b68ed5d24c4","added_by":"auto","created_at":"2026-03-11 12:09:41","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":540349,"visible":true,"origin":"","legend":"\u003cp\u003eMyricetin nanoparticle a) TEM b) SEM\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-8680908/v1/57f95a2bfa7589e372b95094.png"},{"id":103772275,"identity":"ba76bd2d-79df-4044-9514-7b4db8b0df75","added_by":"auto","created_at":"2026-03-02 17:32:29","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":190700,"visible":true,"origin":"","legend":"\u003cp\u003eZeta potential of myricetin nanoparticle\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-8680908/v1/56d4aaa88f05899466f0cbf7.png"},{"id":104400651,"identity":"94b3f438-34b2-487e-9bb2-403b380fe01e","added_by":"auto","created_at":"2026-03-11 12:10:37","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":492596,"visible":true,"origin":"","legend":"\u003cp\u003ea) smd-Myricetin 2D b) smd Myricetin 3D structures\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-8680908/v1/609db480c963d803257cefd5.png"},{"id":104400381,"identity":"20c123dd-e07e-4c9f-ba14-988a66d91074","added_by":"auto","created_at":"2026-03-11 12:09:51","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":453313,"visible":true,"origin":"","legend":"\u003cp\u003ea)1OSE-Myricetin 2D b) Myricetin 3D structures\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-8680908/v1/712152f7b741dedf7afbcd1e.png"},{"id":103772281,"identity":"8632e2c8-cccd-4419-bc78-341f8294c4ce","added_by":"auto","created_at":"2026-03-02 17:32:29","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":44931,"visible":true,"origin":"","legend":"\u003cp\u003eGraphical representation of\u003cstrong\u003e \u003c/strong\u003esum of the glucose levels\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-8680908/v1/2528ca059c42f68bda2d4165.png"},{"id":104400446,"identity":"40a44735-b9db-4349-a610-1cc3ae986592","added_by":"auto","created_at":"2026-03-11 12:10:00","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":407017,"visible":true,"origin":"","legend":"\u003cp\u003eBlood glucose level in diabetic Wistar rat\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-8680908/v1/6148d390503ebd1b545ba7bb.png"},{"id":104780017,"identity":"775bf41b-7899-4dc9-9fbf-3a0bcb843649","added_by":"auto","created_at":"2026-03-17 07:49:16","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4130343,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8680908/v1/49bee4e6-c749-4f87-af8a-8de3275986f6.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Alpha-Amylase Inhibition by Myricetin Nanoparticles: An in Silico, In Vitro, and In Vivo Evaluation toward Natural Antidiabetic Therapy","fulltext":[{"header":"Introduction","content":"\u003cp\u003eDiabetes mellitus, an endocrine disorder characterized by metabolic dysregulation and high blood sugar levels, presents symptoms such as the excessive urination (polyuria); smell of acetone on the patient's breath; rapid, deep breathing (Kussmaul breathing); excessive thirst (polydipsia); excessive hunger (polyphagia); nausea; vomiting; abdominal pain; blurred vision; weight loss; and various altered states of consciousness or arousal (including hostility, mania, confusion, and lethargy). This condition poses perils such as cardiovascular diseases, neuropathy, nephropathy, and retinopathy [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Type I, Type II, gestational, maturity onset at a young age, and malnutrition-related are the types of diabetes[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].Several factors attribute to the development of diabetes, viz, impaired responsiveness to insulin by body\u0026rsquo;s cells, reduced glucose uptake, impaired insulin secretion owing to dysfunction, destruction of pancreatic beta cells, scarcity in insulin production, glucose imbalance, other attributing factors like genetics, obesity, scarcity of physical activity, unhealthy diet, and age[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Regular exercise, a healthy diet, weight loss, and stress management avert diabetes[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. If left untreated, it leads to perils, viz., heart ailments, stroke, peripheral artery disease, kidney damage (even failure may occur), nerve damage leading to numbness, tingling, pain, eye damage, encompassing blindness, and impediments in hearing. Complications of diabetes like cancer, end stage kidney disease ,peripheral artery disease found globally. The high rate of mortality is due to macrovasular and microvascular complications which affects quality of life [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. The increased population that is at risk of developing diabetes needs affordable and effective mitigation strategies [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].Despite the fact that synthetic drugs continue to play a pivotal role in the mitigation of diabetes, compounds that originate from plants have allured attention as a new and promising therapeutic agent owing to their low toxicity and side effects [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].Flavonoids are polyphenolic compounds that are found in abundance in fruits, vegetables, and beverages like tea and wine [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] .They are classified in various subgroups, which are flavonols, flavones, flavanones, and isoflavones[\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Myricetin is a flavonol,[\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e] which has largely been studied with respect to its numerous biological actions. Myricetin is an antioxidant,[\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e] anti-inflammatory,[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e] antimicrobial,[\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] and anticancer drug[\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] and thus a very strong pharmaceutical substance in medications for metabolic and degenerative disease, which include diabetes. Myricetin, a flavonoid found in various plants, has been extensively studied for its antioxidant, neuroprotective,[\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e] and pharmacological properties in vitro. Myricetin is a possible therapy option for neurological ailments based on research revealing it can suppress oxidative stress, boost natural antioxidant enzymes, and neutralize free radicals. Flavonoid compounds, including myricetin, generally possess low bioavailability, which may limit their clinical usefulness despite their established high biological activity. Glycosylation, metabolism, and absorption are factors that affect systemic availability [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. However, evidence exists to demonstrate that dietary flavonoid is safe in dietary quantities[\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Antioxidant and anti-inflammatory activities are significant in averting dysfunction of β-cell and insulin resistance, rather than causing them. Myricetin is common flavonoid available in plants. It is an attenuator of oxidative stress that removes free radicals and amplifies endogenous antioxidant enzymes. Inhibition of α-Glucosidase and α-Amylase Myricetin and other flavonoids prevent the rise of postprandial glucose levels by inhibiting carbohydrate-searching enzymes. This process can be likened to artificial pills such as acarbose. According to the Insulin Mimetic and Secretagogue Action Studies, myricetin boosts glucose intake in an insulin-like way that involves mimicking and hierarchical insulin regulation. It also activates also insulin secretion of pancreatic β-Cell. The neuroprotective potency of myricetin in β-cells of the pancreas has been established through its antioxidative activity which decreases apoptosis and maintains functionality. A few animal experiments elucidate the antidiabetic effect of myricetin. Preclinical studies has shown antihypertensive,antidiabetic ,anticancer and antioxidant activities. [29 ].\u003c/p\u003e \u003cp\u003eTreatments with myricetin enhanced glycemic control and improved the lipid profiles and sensitivity of insulin in diabetic rat. There is some human clinical evidence that supports possible use of polyphenol-rich diets in glycemic control albeit limited. Other Flavonoids Quercetin and the structural analogs luteolin and kaempferol have been able to show antidiabetic effect through similar means, i.e., by enzyme inhibition and anti-inflammatory action. Poor solubility, potency, low bioavailability and the failure to investigate in large scale clinical trials are some of the challenges encountered that may be solved in near future. To address these limitations, nanoparticle delivery systems,[\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e] and glycoside derivatives are under investigation. AutoDock Vina,[\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e] PyRx,[\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e] and Schrodinger Glide[\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e] are employed to run the molecular docking which simulates the small molecule (ligand)-target protein interaction to predict the binding affinity and orientation in the active site. In general, these tools treat the protein receptor as a rigid body, but semi-flexible docking is sometimes permitted, whereas the ligand is assumed to be fully flexible thereby permitting investigation of all rotatable bonds and torsional conformations. The generated poses are scored with a semi-empirical scoring function which estimates binding affinity which is usually reported in kcal/mol, the lower the value of binding energy indicates the higher binding affinity and preferred ligand-receptor interactions, such as hydrogen bonding, van der Waals force, pi pi-stacking and electrostatic interaction, which are usually visualized with a molecular visualization package including Discovery Studio, PyMOL or Chimera. The active site is bounded by a grid box to cut the search space and enhance the docking efficiency. This grid restricts the docking algorithm to a pertinent region on the receptor that increases specificity. The present investigation focuses on assessing the antidiabetic potential of myricetin utilizing both computer-based simulations (\u003cem\u003ein silico\u003c/em\u003e) and laboratory experiments \u003cem\u003e(in vitro\u003c/em\u003e).\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e\u003cstrong\u003eMaterials:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIn silico tools:\u0026nbsp;\u003c/strong\u003eVarious molecular modeling open-source tools such as Discovery Studio, PyMOL or Chimera, Auto dock, \u003csup\u003e\u003cspan lang=\"EN-US\"\u003e[\u003c/span\u003e35\u003cspan lang=\"EN-US\"\u003e,\u003c/span\u003e36\u0026ndash;38\u003cspan lang=\"EN-US\"\u003e]\u003c/span\u003e\u003c/sup\u003e PyRx,\u003csup\u003e\u003cspan lang=\"EN-US\"\u003e[\u003c/span\u003e33\u003cspan lang=\"EN-US\"\u003e]\u003c/span\u003e\u003c/sup\u003e and Schrodinger Glide\u0026nbsp;[39\u0026ndash;43\u003cspan lang=\"EN-US\"\u003e]\u003c/span\u003e etc. were employed in present investigation.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAssay chemicals:\u0026nbsp;\u003c/strong\u003eMyricetin (\u0026ge;98% purity) was purchased from Yucca Enterprises Mumbai, Porcine pancreatic \u0026alpha;-amylase, and Phosphate buffer pH 6.8 were procured from Loba Chemie; Soluble starch from Loba Chemie; All solvents and chemicals were all analytical grades. Deionized water was employed throughout the study. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eChemicals:\u0026nbsp;\u003c/strong\u003eCopper chloride was purchased from Loba Chemie.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003emyricetin\u0026nbsp;nanoparticles\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e0.1 M Copper sulphate dissolved in 100 ml distilled water; stirred the solution to obtain a clear, blue-coloured copper salt solution. Myricetin was slowly added to copper sulphate solution with constant stirring. The mixture was heated at 60\u0026ndash;80 \u0026deg;C for 30\u0026ndash;60 minutes. Colour change typically from blue to dark brown, was noted indicating nanoparticle formation. Reaction mixture was made to stand for a few hours for complete reduction. Mixture was centrifuged at 8000\u0026ndash;10000 rpm for about 15\u0026ndash;20 minutes. The purified nanoparticles dried in a hot air oven at 50-60 \u0026deg;C.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCharacterization\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eof\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003emyricetin\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003enanoparticles\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePhysiochemical studies\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePercent yield of the synthesized derivative, melting point, pH (assess compatibility with biological system), solubility and dispersibility (assess aqueous solubility along with dispersion behaviour) were recorded [44,45].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFTIR\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSamples were analysed by FTIR spectropho\u0026shy;tometer (Alpha T Bruker) in the\u0026nbsp;standard region 4000\u0026ndash;400 cm\u003csup\u003e1\u003c/sup\u003e by potassium bromide method to confirm functional groups, chemical interaction between adjuvants, and nanoparticle components. FTIR spectra were recorded for myricetin\u0026nbsp;nanoparticles. Scans were interpreted for the retention of principle peaks, shifting and new peaks appearance. Figure 1 illustrates FTIR spectra\u0026nbsp;[46,47].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eParticle size\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSEM of was assessed with the help of scanning electron microscope (VEG A3 TESCAN), at 15 keV accelerating voltage. 2 mg samples were coated in vacuum employing thin gold layer before assessment [ 47,48] \u0026nbsp;SEM photographs of myricetin nanoparticles is illustrated in Figure 4.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eZeta potential assessment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eZeta potential assesses surface charge. Zeta potential was recorded for CMGG and nanoparticles employing a zeta potential analyser (Zetasizer Horiba Scientific, Japan). Figure 3 illustrates zeta potential analysis [48].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIn silico molecular docking assessment\u003c/strong\u003e\u003csup\u003e\u003cspan lang=\"EN-US\"\u003e[\u003c/span\u003e49\u003cspan lang=\"EN-US\"\u003e].\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eLigand preparation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e3D structure of myricetin was retrieved from PubChem database (CID: 5281672) in SDF format and converted to PDB format employing Open Babel. Ligand was energy-minimized and converted to PDBQT format employing AutoDock Tools, assigning Gasteiger charges and adding rotatable bonds\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eProtein preparation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCrystal structures of target proteins-human pancreatic \u0026alpha;-amylase (PDB ID: 1HNY) and \u0026alpha;-amylase-acarbose complex (PDB ID: 1OSE) were downloaded from the RCSB Protein Data Bank. Lligands, water molecules, and ions were removed. Polar hydrogens were added, and proteins were converted to PDBQT format.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInteraction analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eLigand-protein interactions were visualized employing Discovery Studio Visualizer 2020. Hydrogen bonding, hydrophobic interactions, and \u0026pi;\u0026ndash;\u0026pi; stacking were assessed to identify key residues in ligand binding within the catalytic site.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIn vitro\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026alpha;\u003c/strong\u003e\u003cstrong\u003e-amylase inhibition assay\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026alpha;-amylase inhibition potential of myricetin and its nanoparticles was determined by Dinitro salicylic acid (DNS) method \u003csup\u003e\u003cspan lang=\"EN-US\"\u003e[\u003c/span\u003e50\u003cspan lang=\"EN-US\"\u003e]\u003c/span\u003e\u003c/sup\u003e. Briefly, 500 \u0026micro;L of test sample was allowed to react with 0.1 M 500 \u0026micro;L of 0.5 % \u0026alpha;-amylase solution (1 U/mL) in Phosphate buffer solution (PBS) pH 6.9 and incubated at 25 \u0026deg;C for 10 min. Subsequently, 500 \u0026micro;L of 1% starch solution in 0.1 M PBS 6.8 was added and incubated at 25 \u0026deg;C for another 10 min. same procedure was redetected for control where 500 \u0026micro;L of the enzyme was replaced by buffer. Reaction was terminated by adding 1000 \u0026micro;L of DNS reagent, followed by boiling at 90 \u0026deg;C for 5 min. Standard acarbose (\u0026alpha;-amylase enzyme inhibitor) was employed as standard drug. After cooling to room temperature, absorbance was measured at 540 nm employing UV spectrophotometer (Lab India 3000). % inhibition of \u0026alpha;-amylase enzyme was computed using formula as follows.\u003c/p\u003e\n\u003cp\u003e\u003cimg src=\"https://myfiles.space/user_files/58895_8739fc6c57c1c19a/58895_custom_files/img1772472290.png\" width=\"785\" height=\"83\"\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStreptozotocin induced antidiabetic activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAs shown in the table no. 1 ,Wistar rats were divided into 5 groups, with 6 rats in each. The second group was Streptozotocin-induced diabetic injection intra peritoneal and did not undergo any treatment. The diabetic of the second group, used as reference, and that of the third to fifth group was treated by an administered orally test item (F1) and Glibenclamide (5 mg/kg), respectively, to elevate their blood glucose levels. Blood samples were collected from the retro orbital of the overnight (12-15h) fasted rats and blood glucose level was determined on 0th, 7th, 14th and 21st day along with body weight and body temperature. If the blood glucose levels of rats\u0026gt; 200 mg/dL then the rats are considered to have hyperglycaemia. In all treated groups, the glucose level was measured before and after the Streptozotocin-induced diabetic injection using a digital glucometer. At specified time intervals (On day 0, 7, 14, 21 after treatment), blood was collected from retro orbital sinuses. Blood glucose levels were determined using a digital glucometer.\u003c/p\u003e"},{"header":"Result and discussion ","content":"\u003cp\u003e\u003cstrong\u003eCharacterization of\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003emyricetin\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003enanoparticles\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe yield of the synthesized\u0026nbsp;myricetin nanoparticles was\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eapproximately in the 75-80%, melting point was 220 \u0026ordm;C, pH was 6.8. Myricetin nanoparticles swelled, formed clear dispersion in water with no signs of precipitation or sedimentation after 24 hours suggesting high solubility. Nanoparticles remained uniformly suspended for long duration with negligible sedimentation thus it can be concluded that nanoparticles exhibited good dispersibility. Thus, it was concluded that synthesized myricetin nanoparticles exhibited desired physical and chemical features. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFTIR\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSamples were analysed by FTIR spectropho\u0026shy;tometer\u0026nbsp;(Alpha T Bruker)\u0026nbsp;in the standard region 4000\u0026ndash;400 cm\u003csup\u003e1\u003c/sup\u003e to confirm functional groups, chemical interaction between myricetin and copper chloride, and nanoparticle components. Figure 1and 2 illustrates FTIR spectra of myricetin and nanoparticles. Hydroxyl and carbonyl groups (\u0026ndash;OH and C=O) present in myricetin and active in binding to metal ions, strong band at 1645 cm⁻\u0026sup1; confirms carbonyl group. Metal\u0026ndash;Oxygen Bond Formation, Peaks at \u0026lt;600 cm⁻\u0026sup1; (e.g., 518, 472, 408 cm⁻\u0026sup1;) represent Cu\u0026ndash;O or Cu\u0026ndash;OH bonds, confirms successful loading or formation of copper nanoparticles with plant-derived compounds. Thus, FTIR spectrum confirms the presence of myricetin functional groups (phenolic \u0026ndash;OH, carbonyl, etc.) Their involvement in the reduction and stabilization of copper nanoparticles. Formation of Cu\u0026ndash;O bonds confirm successful synthesis of myricetin copper nanoparticles through green synthesis routes. \u0026nbsp;Shift in C=O stretch (~1656 \u0026rarr; 1645 cm⁻\u0026sup1;) shows coordination of Cu\u0026sup2;⁺ with carbonyl oxygen in myricetin. Reduction/shift of OH Stretch (~3291 \u0026amp; 2920 cm⁻\u0026sup1;) indicates hydrogen bonding changes or involvement in metal complexation. Appearance of Cu-O peaks (\u0026lt; 600 cm⁻\u0026sup1;) at 677, 518, 472, 408 cm⁻\u0026sup1; are absent in pure myricetin, present only in copper nanoparticles spectrum. Thus, it can be concluded from FTIR that there is successful metal-ligand bond formation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eParticle size\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eParticle size assessment was employed to determine average size, particle size distribution, homogeneity of nanoparticles and was analysed employing a particle size analyser (Figure 3).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe main peak is between 100 and ~300 nm, with most particles clustering around 183\u0026ndash;210 nm. Distribution type was single peak and moderate spread. Polydispersity index was 0.58 indicates a broad size distribution, likely due to aggregation. TEM images of myricetin nanoparticle demonstrate spherical shape and are dense nature with clear boundaries with no aggregation. Nanoparticle demonstrates 30-40 nm in diameter; thus, it was concluded that these nanoparticles were in nanometre range. SEM of myricetin nanoparticle also demonstrate its spherical nature. Figure 4a and 3b illustrates TEM and SEM of myricetin nanoparticle respectively.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eZeta potential assessment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eZeta potential assesses surface charge. Zeta potential was recorded for myricetin nanoparticles employing a zeta potential analyser and was found to be -35 mV hold a moderately negative surface charge and nanoparticles are stable as zeta potential value is closer to \u0026plusmn;30 mV. Figure 5 illustrates zeta potential analysis.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.1 Molecular docking\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMolecular docking analysis has been performed against two critical diabetes-related proteins \u0026alpha;-amylase and 1OSE employing AutoDock, PyRx, and Schrodinger Glide. The least binding affinity represents the strongest binding affinity. Binding affinity (kcal/mol) indicates strength of ligand binding; more negative values suggest stronger binding. Root Mean Square Deviation, upper bound (RMSD/ub) measures deviation from the reference pose. RMSD/lb (lower bound) like ub, typically employed to validate docking accuracy. Lower RMSD values (\u0026lt;2 \u0026Aring;) indicate more reliable binding poses. \u0026nbsp;Figure 6a demonstrates smd-Myricetin 2D and 6b smd Myricetin 3D structures respectively. Figure 7a demonstrates 1OSE-Myricetin 2D and 7b Myricetin 3D structures respectively. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMyricetin binds effectively to 1OSE with multiple stable conformations; docking is robust, with several poses under 2 \u0026Aring; RMSD supporting pose stability and reproducibility. Molecular docking results of myricetin against 1OSE protein revealed a highly favourable interaction profile. Top-ranked pose exhibited a binding affinity of -8.8 kcal/mol, suggesting a strong and energetically stable interaction between ligand and protein\u0026rsquo;s active site. This value reflects a high probability of effective binding under physiological conditions, making myricetin a promising candidate for modulating 1OSE-related activity. Root mean square deviation (RMSD) of this top pose was 0 \u0026Aring;, confirming that it is reference conformation-the most accurate and least deviated pose generated during the docking simulation. RMSD of 0 implies that this pose was used as a baseline for comparing other generated conformations and likely represents the most native-like fit within the binding pocket. Several additional poses demonstrated RMSD values less than 2 \u0026Aring; (e.g., 2.812 \u0026Aring;, 1.690 \u0026Aring;, 1.275 \u0026Aring;), which are within accepted range for reliable docking predictions. Poses with RMSD \u0026lt; 2 \u0026Aring; are generally considered to maintain a similar binding orientation as the reference pose, indicating high reproducibility and consistency of ligand binding across docking iterations. These conformations further validate the robustness of docking protocol and suggest that Myricetin can adopt multiple stable binding geometries in 1OSE active site. In contrast, several poses exhibited significantly higher RMSD values (e.g., 17.073 \u0026Aring;, 17.815 \u0026Aring;, 8.99 \u0026Aring;), suggesting these conformations likely represent alternative binding modes or non-optimal orientations within or near binding cavity. myricetin, a naturally occurring flavonol, exhibits a strong and stable binding affinity toward \u0026alpha;-amylase, a key enzyme involved in carbohydrate digestion and postprandial hyperglycaemia. Molecular docking results revealed a highly favourable binding energy, indicating that myricetin can effectively occupy the active site of \u0026alpha;-amylase and potentially inhibit its enzymatic activity. Detailed interaction analysis showed that myricetin forms multiple hydrogen bonds and hydrophobic contacts with critical residues within the catalytic pocket, suggesting competitive inhibition as a likely mechanism of action.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIn vitro\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026alpha;\u003c/strong\u003e\u003cstrong\u003e-amylase inhibition assay\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe digestive enzyme \u0026alpha;-amylase is responsible for hydrolysing dietary starch (maltose), which breaks down into glucose prior to absorption. Inhibition of \u0026alpha;-amylase can lead to reduction in post prandial hyperglycaemia in diabetic condition. Given sample demonstrated good inhibition of \u0026alpha;-amylase enzyme when compared to acarbose standard.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStreptozotocin induced antidiabetic activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe results were expressed as mean SD (n=6), ns p\u0026gt;0.05, non- significant; p \u0026lt;0.01, When compared with positive control group. Based on the provide data on glucose levels over the course of the experiment, here is a conclusion drawn regarding the anti-diabetic activity.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e Boold glucose level in diabetic Wistar rat\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGlucose\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003elevel on day\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0 (mg/dl)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGlucose\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;level on day\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e7 (mg/dl)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGlucose\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003elevel on day\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e14 (mg/dl)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGlucose\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003elevel on day\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e21(mg/dl)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eNormal group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e85.98\u0026plusmn;4.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e88.45\u0026plusmn;2.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e86.36\u0026plusmn;3.69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e85.25\u0026plusmn;4.28\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eDiabetic control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e230.97\u0026plusmn;1.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e220.24\u0026plusmn;4.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e275.28\u0026plusmn;5.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e270.18\u0026plusmn;7.28\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eStandard\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e225.3\u0026plusmn;4.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e258.57\u0026plusmn;2.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e160.51\u0026plusmn;0.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e110.68\u0026plusmn;0.98\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eNanoparticles\u003c/p\u003e\n \u003cp\u003emyricetin (100 mg/kg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e227.58\u0026plusmn;1.36\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e270.18\u0026plusmn;4.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e150.18\u0026plusmn;7.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e95.35\u0026plusmn;5.20\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eBlood glucose lowering study (hypoglycemic study)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMyricetin nanoparticles showed a significant and consistent reduction in blood glucose levels over 21 days (figure 8) After an initial rise by day 7 due to induced hyperglycaemia, treatment led to a marked decrease by day 14 and near-normal levels by day 21. This improved effect is attributed to enhanced bioavailability and targeted delivery through the nanoparticle formulation. Compared to the simple extract and Glibenclamide, Myricetin nanoparticles demonstrated superior anti-diabetic activity, likely due to their antioxidant and insulin-sensitizing properties. These findings support the potential of Myricetin nanoparticles as an effective anti-diabetic agent. Figure 9 illustrates blood glucose level in diabetic Wistar rat.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe current investigation infers that myricetin nanoparticles exhibited significant α-amylase inhibitory potential, emphasizing their propensity as a natural antidiabetic entity. Molecular docking assessment revealed strong binding affinity of myricetin to the active site of α-amylase, assisted by favourable interaction energies and key hydrogen bond formations. In vitro α-amylase inhibition assay further supported outcome of the current investigation, with myricetin nanoparticles exhibiting ameliorated inhibition. The blend in silico and in vitro approach highlights the therapeutic promise of myricetin-loaded nanoparticles in mitigating hyperglycemia, presenting a cost-effective and substitute to conventional therapies.\u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eConflict of Interest:\u003c/h2\u003e \u003cp\u003eThe authors declare no conflict of interest\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding:\u003c/h2\u003e \u003cp\u003eThe authors did not receive support from any organization for the submitted work.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eKarishma Ramchandra Waghmare: Conceptualization, formal analysis, experimental, methodology, data curation and writing original draft; Guno Sindhu Chakraborthy: Resources, and literature survey; supervision, review and editing.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThe authors are grateful to Yucca Enterprises, Mumbai, India, for providing Myricetin used as a marker in this study. The authors sincerely thank ICON LABS, ICON HOUSE, India, and Yashwantrao Chavan Institute of Science, Satara, for their valuable support in carrying out the characterization studies. The authors also extend their gratitude to Invitox R and D Institute, Pune, India, for conducting the in vivo activity. The generous laboratory facilities provided by the Principal, Shri DD Vispute college of pharmacy and Research center New Panvel, India, gratefully acknowledged.Data Availability All data generated or analysed during this study are included in this published article.DeclarationsConflict of Interest : The authors declare no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAntar SA, Ashour NA, Sharaky M, et al. Diabetes mellitus: Classification, mediators, and complications; A gate to identify potential targets for the development of new effective treatments. 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Bhowate. Dinesh R. Chaple DrAJ asnani, PI, AVLVSB. Molecular Docking: A Powerful Tool In Modern Drug Discovery And Its \u0026nbsp;Approaches .\u003c/li\u003e\n \u003cli\u003eIbrahim MM, Zaki ER, Rady MR. Alpha-amylase inhibitory activity and in silico studies of in vitro sweet basil plantlets treated with chitosan and ZnO NPs. In Vitro Cellular \u0026amp; Developmental Biology - Plant. 2024;60(2):147-160. doi:10.1007/s11627-023-10401-0\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Alpha-amylase, diabetes, in silico, in vitro, molecular docking, myricetin","lastPublishedDoi":"10.21203/rs.3.rs-8680908/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8680908/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003ePurpose\u003c/h2\u003e \u003cp\u003eDiabetes mellitus is a worldwide health challenge, concern, and effective and affordably priced therapies are pivotal. Myricetin, a naturally occurring flavanol, has demonstrated promising antidiabetic potential. The current research aims to investigate the binding affinity and molecular interactions of myricetin with alpha-amylase and 1OSE protein, and to explore its potential as a therapeutic agent for diabetes management.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eThe present investigation, allied with molecular docking of myricetin against diabetes related targets, was investigated through Auto dock Vina, PyRx, and Schrodinger Glide. The target proteins were prepared in PDBQT format. Myricetin was docked to explore its molecular mechanism. The binding affinity and orientation of myricetin in the active site of alpha-amylase and 1OSE protein were evaluated. The in vivo antidiabetic potential of myricetin nanoparticles was assessed in Streptozotocin-induced diabetic Wistar rats. The experimental protocol was approved by the Institutional Animal Ethics Committee (IAEC) of Invitox R and D institute registration no.2273/PO/RERC/S/23 CCSEA, Report no. IRDI/IVAS/18/2024-25 registered under CPCSEA. The animals were randomly assigned to five groups (n\u0026thinsp;=\u0026thinsp;6). Group I served as the normal control, while Group II received intraperitoneal Streptozotocin and was maintained as the untreated diabetic control. Groups III and IV were administered the test formulation (F1) orally at the selected doses, whereas Group V received the standard antidiabetic drug glibenclamide (5 mg/kg). The F1-treated groups exhibited a significant decrease in fasting blood levels of glucose compared with the diabetic control group, confirming in vivo antidiabetic efficacy of myricetin nanoparticles.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eMyricetin exhibited a strong and stable binding affinity toward alpha-amylase and 1OSE protein, with a binding energy of -8.8 to -7.6 kcal/mol. Molecular docking results revealed multiple hydrogen bonds and hydrophobic contacts between myricetin and critical residues within the catalytic pocket.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eMyricetin exhibits promise as a therapeutic propensity for diabetes mitigation owing to its strong binding affinity and inhibitory activity against alpha-amylase.\u003c/p\u003e","manuscriptTitle":"Alpha-Amylase Inhibition by Myricetin Nanoparticles: An in Silico, In Vitro, and In Vivo Evaluation toward Natural Antidiabetic Therapy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-02 17:32:24","doi":"10.21203/rs.3.rs-8680908/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"cdfbed7e-f0d6-4175-82d4-ed4332f77850","owner":[],"postedDate":"March 2nd, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-04-12T03:08:51+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-02 17:32:24","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8680908","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"identity":"rs-8680908","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}