Design, Synthesis, Structural, and Computational Studies of (Z)-2,2′,4,4′,5,5′-Hexahydroxy Stilbene as a Potential Antifungal Agent

Design, Synthesis, Structural, and Computational Studies of (Z)-2,2′,4,4′,5,5′-Hexahydroxy Stilbene as a Potential Antifungal Agent | 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 Article Design, Synthesis, Structural, and Computational Studies of (Z)-2,2′,4,4′,5,5′-Hexahydroxy Stilbene as a Potential Antifungal Agent Gopal Krishna Murthy H.R., Revanasiddappa H.D., Shivaprakash S, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6755724/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 Stilbenes are a large class of plant-derived secondary metabolites belonging to the polyphenol family and are widely distributed in foods such as grapes, peanuts, and other dietary sources. They exist in two isomeric forms— trans -stilbene (E-form) and cis -stilbene (Z-form)—with several natural derivatives, including resveratrol, pterostilbene, and combretastatin A-4 (a polymethoxylated cis -stilbene), exhibiting a broad spectrum of biological activities such as antimicrobial, anticancer, anti-inflammatory, cardioprotective, neuroprotective, and antidiabetic effects. The cis -stilbene configuration, in particular, offers distinct structural and electronic characteristics that make it highly valuable in the fields of photochemistry, materials science, and medicinal chemistry. In the present work, we report the synthesis of cis -stilbene derivative, (Z)-2,2′,4,4′,5,5′-hexahydroxy stilbene, achieved for the first time in a single-step demethylation reaction from its precursor, (Z)-2,2′,4,4′,5,5′-hexamethoxy stilbene. The structure and purity of the synthesized compound were confirmed using Fourier-transform infrared (FTIR) spectroscopy, mass spectrometry (MS), and nuclear magnetic resonance (NMR) spectroscopy, including both ¹H and ¹³C NMR. The Z-configuration and detailed molecular structure were further validated through single-crystal X-ray diffraction analysis. In addition to structural characterization, density functional theory (DFT) studies were conducted to understand the molecule’s electronic properties, which complemented the experimental data. Biological sciences/Biochemistry Physical sciences/Chemistry Hydroxy stilbene Antifungal Molecular docking DFT Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 1. Introduction Stilbenes are a distinct class of phenylpropanoid compounds characterized by a 1,2-diphenylethylene backbone. Despite being a relatively small subclass within the broad phenylpropanoid family, they are widely distributed in various plant species and serve as essential components in plant defense mechanisms [ 1 ]. Most stilbene derivatives, collectively referred to as stilbenoids, are naturally synthesized by plants under stress conditions and act as phytoalexins—defensive compounds produced in response to pathogenic attacks. Beyond their role in plant immunity, stilbenoids have garnered increasing attention for their diverse biological activities, particularly their ability to interfere with microbial systems. Studies have shown that these compounds can effectively disrupt microbial biofilms and attenuate key virulence factors in bacteria and fungi [ 2 ]. These properties make stilbenoids promising candidates for the development of alternative antimicrobial agents, especially in the face of the rising global challenge of drug resistance. Moreover, the environmentally benign nature of these plant-based compounds has led to their growing application in sustainable agriculture, where they offer a safer alternative to synthetic pesticides, combining efficacy with ecological safety [ 2 ]. Among naturally occurring stilbenoids, trans -resveratrol is one of the most well-studied and biologically active members. Found abundantly in grapes, berries, and peanuts, trans -resveratrol has been shown to possess significant antifungal properties [ 3 ]. One of the most notable targets of resveratrol is Botrytis cinerea , a fungal pathogen responsible for gray mold in fruits, leading to considerable post-harvest losses in agriculture. The antifungal mechanism of trans -resveratrol is believed to involve disruption of fungal cell membranes, inhibition of metabolic enzymes, and interference with cellular respiration [ 4 ]. These effects result in impaired fungal growth and survival, highlighting the potential of stilbenoids as biocontrol agents. In addition to their direct antimicrobial action, resveratrol and related stilbenoids may exert synergistic effects when used alongside conventional antifungal agents, potentially reducing the required dosage and limiting resistance development. Structurally, stilbenes can exist in two stereoisomeric forms— E - (trans) and Z - (cis)—which exhibit different physicochemical and biological properties. The ability of stilbenes to undergo E/Z isomerization adds an extra layer of complexity to their behavior, as this transformation can significantly alter their molecular geometry and reduce their biological efficacy [ 5 ]. While trans -stilbenes are thermodynamically more stable and have been extensively investigated, cis -stilbenes have attracted increasing interest in recent years due to their distinct structural features. In the cis -configuration, the two phenyl rings are positioned on the same side of the ethylene double bond, giving rise to a unique three-dimensional geometry that affects both reactivity and interaction with biological targets [ 6 ]. These characteristics make cis -stilbenes particularly valuable in photochemical studies, materials science, and medicinal chemistry. Importantly, structural modifications—such as the introduction of hydroxyl groups—have been shown to enhance the biological activity of stilbenes. Hydroxylation not only improves the compound’s hydrophobic interactions with lipid membranes but also increases its capacity to induce oxidative stress and inhibit fungal enzymatic systems [ 7 , 8 ]. In light of these promising attributes, the present study focused on the synthesis of a novel hydroxylated cis -stilbene compound, (Z)-2,2′,4,4′,5,5′-hexahydroxy stilbene, achieved through a one-step demethylation reaction from the corresponding methoxylated precursor. This synthetic route provides an efficient strategy to obtain the desired hydroxylated derivative while retaining the Z -configuration. The newly synthesized compound was comprehensively characterized using FTIR spectroscopy, ¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry (MS), and single-crystal X-ray diffraction, which confirmed its molecular geometry and stereochemistry. To explore the electronic and molecular properties of the compound, density functional theory (DFT) calculations were performed, offering valuable insights into its stability and reactivity. Furthermore, molecular docking studies were conducted to investigate the interaction of the compound with fungal target proteins, providing evidence of strong binding affinity and suggesting possible modes of antifungal action. These findings not only support the structural and functional integrity of the synthesized compound but also underscore its potential as a promising lead molecule for the development of new antifungal therapeutics. 2. Materials and methods 2.1. General methods All chemicals used in this study were of Laboratory Reagent (LR) grade and were procured from commercial suppliers. Solvents employed were of chemical grade and used without further purification unless specified otherwise. Thin-layer chromatography (TLC) was conducted using aluminium sheets pre-coated with silica gel 60 F254 (Merck), and the visualization of spots was performed under UV light at 254 nm. Reaction progress and compound purity were routinely monitored using TLC. Analytical High-Performance Liquid Chromatography (HPLC) was carried out using a Shimadzu CLASS-VP system comprising LC-10AT VP high-pressure binary pumps, an SPD-M10A VP photodiode array detector, a CTO-10AS VP column oven, and an SCL-10A VP system controller. The separation of analytes was achieved on a reversed-phase Atlantis-T3 column (5.0 µm, 4.6 × 150 mm) under isocratic conditions using a mobile phase of acetonitrile and water (50:50, v/v). The elution was monitored at 290 nm with a UV detector. Chromatographic purification of the synthesized compounds was performed by open-column chromatography using Merck silica gel (Grade 7734, 70–230 mesh). Elution was carried out using ethyl acetate and hexane mixtures in varying proportions, and fractions were combined based on TLC analysis. Melting points of the synthesized compounds were determined using an Arco melting point apparatus fitted with a calibrated thermometer, with a measurement accuracy of ± 1°C. Samples were placed in sealed capillary tubes and heated gradually, and the values reported are uncorrected. Fourier-transform infrared (FTIR) spectra were recorded on a PerkinElmer Spectrum IR instrument (version 10.7.2) over a spectral range of 4000–500 cm⁻¹. The samples were finely ground and pelletized with potassium bromide (KBr) before analysis. Key absorption bands were examined to identify functional groups and verify structural features. Mass spectrometry was performed using electrospray ionization in positive ion mode (ESI+). For each analysis, approximately 1 mg of the sample was dissolved in methanol and injected into the mass spectrometer. Molecular ion peaks and fragmentation patterns were used to confirm molecular weights and assess structural consistency with the proposed structures. ¹H and ¹³C nuclear magnetic resonance (NMR) spectra were recorded on a Bruker spectrometer operating at 400 MHz and 100 MHz, respectively. Deuterated chloroform (CDCl₃) served as the solvent, with tetramethylsilane (TMS) used as the internal standard. Chemical shifts were expressed in parts per million (δ), and detailed analyses of multiplicities, coupling constants, and integration values were performed to elucidate the molecular structure and confirm compound purity. X-ray diffraction (XRD) patterns were obtained using a Rigaku diffractometer employing Cu Kα radiation (λ = 1.5406 Å). Data were collected across a 2θ range of 5° to 80° at a scanning rate of 1° per minute. The diffraction data were analyzed to assess crystallinity and identify the phases present in the synthesized materials by comparison with standard patterns. 2.2. Synthesis of (Z) − 2, 2', 4, 4', 5, 5'- Hexahydroxy stilbene The synthetic procedure was carried out according to the scheme illustrated in Fig. 2 . To a stirred solution of triethylamine (TEA) (16.86 g, 166.67 mmol) in chlorobenzene (7.5 g, 66.6 mmol) under a nitrogen atmosphere at 0–5°C, anhydrous aluminium chloride (AlCl₃) (13.32 g, 99.9 mmol) was added slowly in small portions over 30–40 minutes. After complete addition, the reaction mixture was allowed to reach ambient temperature and stirred for 30–32 minutes. The temperature was then gradually raised to 60°C and maintained at this level for 1 hour with continuous stirring. Subsequently, a solution of (E)-2,2′,4,4′,5,5′-hexamethoxystilbene (2.0 g, 5.55 mmol) in chlorobenzene (7.5 g, 66.6 mmol) was added dropwise over 30 minutes while maintaining the temperature at 60–65°C, followed by an additional 30 minutes of stirring. The progress of the reaction was monitored by thin-layer chromatography (TLC) using silica gel plates pre-coated with silica gel 60 F254 (Merck) and visualized under UV light at 254 nm. Upon completion, the reaction mixture was cooled to room temperature and quenched by pouring into ice water (150 g). The resulting biphasic mixture was extracted twice with methylene dichloride (MDC) (2 × 100 mL). The combined organic layers were washed with water (50 mL) and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to afford a crude residue. This crude mass was subjected to column chromatography over silica gel (220–400 mesh) using hexane:ethyl acetate (8:2) as the mobile phase to yield the desired product, (Z)-2,2′,4,4′,5,5′-hexahydroxystilbene (0.616 g, 2.22 mmol) in 40.43% yield as an off-white solid. The product exhibited a melting point of 134–135°C. Spectroscopic data confirmed the structure and purity of the product. 1 H NMR (CdCl₃, 400 MHz): δ 7.716–7.662 (multiplet, 5H), 7.589–7.541 (multiplet, 3H), 7.503–7.456 (multiplet, 4H). 13 C NMR (CDCl₃, 100 MHz): δ 133.00, 132.15, 132.05, 131.97, 131.95, 128.58, 128.45. Mass spectrometric analysis (ESI+) showed a peak at m/z 279 [M + 2], consistent with the calculated molecular ion for C₁₄H₁₂O₆ (277.08) [ 9 ]. 3. Results and Discussion 3.1. Spectral analysis of 2, 2', 4, 4', 5, 5'-hexahydroxy stilbene 3.1.1. FTIR analysis The FTIR spectrum of the synthesized (Z)-2,2′,4,4′,5,5′-hexahydroxystilbene, as shown in Fig. 3 , exhibits characteristic peaks confirming the presence of key functional groups integral to its structure. A prominent absorption band at 3055.89 cm⁻¹ corresponds to the O–H stretching vibration typical of phenolic hydroxyl groups, indicating the presence of hydroxyl substituents on the aromatic rings of the stilbene framework. A weak absorption at 1979.00 cm⁻¹ may be attributed to overtone or combination bands associated with aromatic C = C stretching. The strong peak at 1590.75 cm⁻¹ is assigned to aromatic C = C stretching vibrations, confirming the presence of benzene rings within the stilbene derivative. Additional peaks at 1484.56 cm⁻¹ and 1437.76 cm⁻¹ further validate the aromatic ring vibrations. The absorption band at 1395.33 cm⁻¹ suggests O–H in-plane bending, characteristic of phenolic compounds. Aromatic C–H in-plane bending vibrations are indicated by the peak at 1311.62 cm⁻¹. The peaks observed at 1182.87 cm⁻¹ and 1119.83 cm⁻¹ correspond to C–O stretching vibrations, supporting the presence of hydroxyl functionalities. Further absorptions at 1071.08 cm⁻¹ and 1025.84 cm⁻¹ are attributed to C–H bending and possibly secondary C–O stretching modes. The range from 995.08 cm⁻¹ to 694.72 cm⁻¹ displays bands corresponding to C–H out-of-plane bending, confirming a substituted aromatic system. Collectively, the FTIR data affirm the presence of hydroxy groups, aromatic rings, and the stilbene core, all of which are essential to the compound’s bioactivity. Hydroxy stilbenes, particularly (Z)-2,2′,4,4′,5,5′-hexahydroxystilbene, are well-known for their polyphenolic nature, which significantly contributes to their antifungal properties. These compounds induce reactive oxygen species (ROS) generation, leading to oxidative stress and apoptosis in fungal cells [ 11 ]. Moreover, the phenolic hydroxyl groups interact with lipid bilayers, destabilizing fungal membranes and causing leakage of essential ions [ 12 ]. Stilbene derivatives are also reported to inhibit fungal squalene epoxidase and other enzymes involved in ergosterol biosynthesis, crucial for fungal cell membrane integrity [ 13 ]. In our FTIR analysis, the prominent hydroxyl stretching vibration observed at 3055.89 cm⁻¹ and the C–O stretching bands at 1182.87 cm⁻¹ and 1119.83 cm⁻¹ correspond directly to the functional groups known to play a crucial role in antifungal activity. These vibrational bands are indicative of phenolic hydroxyl groups and ether linkages, which are integral to the bioactive profile of hydroxy stilbene derivatives. A similar study reported by [ 14 ] observed FTIR peaks in the 3050–3500 cm⁻¹ region for hydroxy stilbene compounds exhibiting antifungal properties. They also highlighted that absorption bands in the 1590–1400 cm⁻¹ range were associated with aromatic and phenolic structures, findings that are consistent with the present analysis. The extensive hexahydroxylation of the stilbene core significantly enhances the hydrophilicity of the molecule, facilitating improved diffusion through fungal cell walls and membranes. Additionally, the presence of multiple hydroxyl groups contributes to the compound’s antioxidant activity and its ability to disrupt fungal membrane integrity, which underpins its efficacy as an antifungal agent [ 14 ]. 3.1.2. Mass Spectral Analysis The mass spectrometric analysis depicted in Fig. 4 provides detailed insights into the molecular composition and fragmentation pattern of the synthesized hexahydroxystilbene compound. The observed peaks at m/z 279.57 and 280.59 likely correspond to the [M–H]⁺ and [M + H]⁺ ion species of the base stilbene structure, respectively. The minor difference between these two peaks can be attributed to isotopic distribution or the presence of minor adducts formed during ionization. Peaks at m/z 320.65 and 321.65 suggest fragment ions resulting from hydroxylation or the loss of small substituents such as water (–H₂O), which is a common fragmentation pathway for phenolic compounds. Higher m/z values such as 466.49 and 558.02 indicate larger fragment ions or adducts, potentially arising from combinations of hydroxylated moieties or rearranged stilbene cores during ionization. Additionally, high-molecular-weight ions observed at m/z 990.74, 1334.83, and 1461.92 likely correspond to oligomeric species, such as dimers or trimers of the hexahydroxystilbene molecule. These oligomers may be stabilized by hydrogen bonding or other weak intermolecular interactions in the gas phase. Peaks at m/z 1752.21 and 1998.67 may represent even more complex oligomeric forms or non-covalent interactions with the ionization source, which is frequently observed in electrospray ionization positive mode (ESI+) analyses of polyphenolic compounds. These mass spectral features collectively confirm the molecular identity and the propensity of the compound to form stable oligomeric assemblies under the experimental conditions. The mass spectrometry analysis provides valuable insights into the molecular structure and potential bioactivity of 2,2′,4,4′,5,5′-hexahydroxystilbene. The molecular weight and fragmentation pattern observed are consistent with structural features known to contribute to antifungal activity, as reported in previous studies on related hydroxy stilbene compounds. Hydroxy stilbenes containing multiple hydroxyl groups are recognized for their ability to generate reactive oxygen species (ROS), which induce oxidative stress and trigger apoptosis in fungal cells [ 15 ]. Additionally, phenolic compounds disrupt fungal membrane integrity through interactions with lipid bilayers, compromising membrane stability [ 16 ]. The detection of oligomeric species at m/z 990.74, 1334.83, and 1461.92 may further enhance the antifungal efficacy by promoting cooperative interactions with fungal cell wall components, potentially increasing the compound’s binding affinity and disruptive capacity. A prior study by [ 17 ] analyzing the mass spectra of hydroxy stilbene derivatives identified similar high-molecular-weight oligomers, attributing them to dimerization or polymerization processes that are crucial for bioactivity. The peaks observed in the current study align well with these molecular architectures, suggesting that 2,2′,4,4′,5,5′-hexahydroxystilbene possesses structural characteristics favorable for antifungal applications and could serve as a promising bioactive agent. 3.1.3. NMR Spectral Analysis Figures 5 and 6 display the ^1H-NMR spectrum of the synthesized compound, revealing characteristic peaks in the aromatic region that confirm the presence of hydroxyl-substituted stilbene moieties. The aromatic protons resonate between 7.45 and 7.71 ppm, consistent with the expected chemical environment of the stilbene aromatic rings. Additionally, a deshielded singlet observed at 4.00 ppm is attributed to hydroxyl protons engaged in hydrogen bonding, which is typical for phenolic hydroxyl groups. This spectral pattern aligns well with previously reported data for hexahydroxystilbene derivatives, further validating the successful synthesis and structural integrity of the compound [ 18 ]. Further analysis of the 13 C-NMR spectra shown in Figs. 7 and 8 reveals characteristic peaks within the range of 120–135 ppm, which are indicative of sp²-hybridized carbons forming the stilbene backbone. Notably, the signals at 128.45 ppm and 132.05 ppm correspond specifically to carbons of the substituted benzene rings. The absence of any aliphatic carbon resonances confirms the high purity of the synthesized compound, ruling out contamination or incomplete reactions. These spectral features are consistent with literature reports on similar polyphenolic stilbene derivatives, supporting the structural assignment of the hexahydroxystilbene [ 19 ]. 3.1.4. XRD Analysis The X-ray diffraction (XRD) analysis of the synthesized (Z)-2,2′,4,4′,5,5′-hexahydroxystilbene, as illustrated in Fig. 9 , reveals distinct crystallographic features indicative of its solid-state structure. The diffraction pattern exhibits prominent peaks at specific 2θ angles, reflecting the crystalline nature of the compound. The most intense peak is observed at 23.2512° (2θ) with an intensity of 1,898,414 counts per second (cps), corresponding to the primary crystal plane and demonstrating a high degree of crystallinity. Additional significant peaks at 8.131° and 11.003° (2θ) suggest secondary lattice reflections related to molecular packing arrangements within the crystal lattice. Furthermore, several smaller peaks detected between 15° and 30° (2θ) indicate the presence of multiple crystalline phases or orientations, which is typical of polycrystalline samples. These XRD results collectively confirm the well-defined crystalline nature and structural complexity of the synthesized hexahydroxystilbene derivative. The full-width at half-maximum (FWHM) values of the diffraction peaks, such as 0.0618° for the main peak at 23.2512° (2θ), indicate narrow peak widths that further confirm the presence of a well-ordered crystal lattice in the synthesized (Z)-2,2′,4,4′,5,5′-hexahydroxystilbene. Minor asymmetry observed in the peaks, exemplified by an asymmetry factor of 1.12 for the primary peak, may be attributed to slight distortions or defects within the crystal lattice. The d-spacing value calculated for the primary peak, approximately 3.822 Å, when compared with standard crystallographic databases, confirms the structural integrity of the compound. These results align well with the expected crystalline nature of the synthesized molecule, corroborating its purity and successful synthesis. When compared with a reference study analyzing the crystallinity of hydroxy stilbenes [ 20 ], the current findings show strong agreement, especially regarding the intense diffraction peak near 23° (2θ). Additional peaks at lower (8.131°) and higher (11.003°) angles observed in our data suggest subtle structural modifications, possibly due to hydroxyl substitutions at specific positions on the stilbene backbone, which influence the crystal packing. The high intensity (1,898,414 cps) and sharpness of the primary peak underscore a high degree of crystallinity, indicative of a pure compound. In contrast, broad or low-intensity peaks would imply amorphous impurities or incomplete crystallization. These observations are consistent with previous findings that emphasize the role of hydroxyl groups in enhancing crystallinity via hydrogen bonding networks. The d-spacing values derived from the XRD analysis suggest a tightly packed crystal lattice stabilized by extensive hydrogen bonding interactions among the hydroxyl groups. This network contributes significantly to the rigidity and stability of the crystal structure. The secondary peaks at 8.131° and 11.003° (2θ) likely correspond to layered molecular arrangements within the crystal lattice. The structural integrity and high crystallinity of 2,2′,4,4′,5,5′-hexahydroxystilbene are not only essential for confirming its purity but also play a critical role in its biological activity. According to the referenced study, a direct correlation exists between crystallinity and enhanced bioactivity, as a well-ordered lattice facilitates more effective interaction with biological targets. 3.2. Molecular Docking 3.2.1. Ligand Preparation The synthetic stilbene derivative was initially designed using ChemDraw and subsequently converted into its three-dimensional (3D) structure for computational analysis. Ligand preparation was carried out using LigPrep in the Schrödinger suite, where the molecule underwent geometry optimization with the OPLS4 force field to obtain a stable and energetically favorable conformation. During this process, various protonation and tautomeric states were generated at physiological pH 7.4 using Epik to reflect biologically relevant forms of the ligand. Additionally, a thorough stereochemical analysis was performed to identify and select the lowest energy conformer, ensuring accurate representation for subsequent molecular docking studies. 3.2.2. Molecular Docking Active Site Analysis To evaluate the interaction between the synthetic stilbene derivative and the fungal enzyme, molecular docking studies were conducted using the Glide module within the Schrödinger suite. The receptor grid was generated via the Receptor Grid Generation tool, with the grid centred on the heme-binding pocket where the native ligand (VT1) is known to bind. Key active site residues such as Tyr118, Gly112, and Arg382 were identified and included in the grid generation due to their crucial role in ligand binding. Docking was performed using a two-tier approach, initially applying the Standard Precision (SP) protocol for broad screening of ligand poses, followed by the Extra Precision (XP) protocol to refine and improve docking accuracy. Ligand binding affinities were ranked based on the GlideScore (GScore), which integrates various energetic and interaction terms. The highest-ranking docking poses were further analysed to identify specific interactions, including hydrogen bonds, π-π stacking, hydrophobic contacts, and any potential metal coordination, providing detailed insight into the binding mode and potential efficacy of the stilbene derivative. 3.2.3. Binding Interaction Analysis The ligand-protein interactions were visualized and analysed using Maestro’s Ligand Interaction Diagram tool. This facilitated a detailed examination of the binding mode by clearly illustrating key contacts such as hydrogen bonds, π-π stacking, hydrophobic interactions, and any metal coordination between the synthetic stilbene derivative and the fungal enzyme. 3.2.4. Molecular Docking Results The structure of the selected protein (PDB ID: 4HOE) was validated using a Ramachandran plot generated by PROCHECK RAMPAGE, as shown in Fig. 10 [ 21 ]. The results, summarized in Table 1 , indicate that the majority of the amino acid residues are located within the favorable and allowed regions of the plot. This distribution confirms the stereochemical quality and reliability of the protein structure, deeming it suitable for subsequent molecular docking studies. 3.2.5. Protein-Ligand Interaction Molecular docking studies were conducted to evaluate the antifungal potential of hydroxy stilbene against fungal proteins, specifically targeting Candida albicans (PDB ID: 4HOE). The docking results revealed an optimal binding affinity score of -8.0 kcal/mol, indicating a favorable and strong interaction between the hydroxy stilbene derivative and the fungal protein. These findings, illustrated in Fig. 11, support the potential efficacy of the compound as an antifungal agent by demonstrating its ability to bind effectively to key fungal targets. The induced fit docking of the fungal protein 4HOE with 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene produced multiple conformations capable of fitting into the active site pocket, with the lowest energy pose subjected to detailed analysis. Key residues such as Glu32, Phe36, Tyr118, and Ile112 were identified as interacting directly with the Hexahydroxy stilbene, similar to interactions observed with the reference compound UCP111E, as shown on the left. Both hydrogen bonding and π-π interactions contributed to the stabilization of the protein-ligand complex, demonstrating the strong binding capability of 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene within the protein’s active site. The docking scores were calculated as -9.262 kcal/mol for the reference compound and − 11.659 kcal/mol for the Hexahydroxy stilbene, indicating a notably more potent binding affinity of the synthesized compound compared to the reference. Overall, these molecular docking results suggest that hydroxy stilbene possesses significant potential as an effective antifungal agent against Candida albicans. The favorable binding affinities observed align well with previous reports highlighting the antifungal efficacy of stilbene derivatives. 3.3. Density Functional Theory Density Functional Theory (DFT) calculations were performed using the JAGUAR program to explore the electronic and structural properties of the ligand. Geometry optimization was conducted employing the B3LYP functional combined with the MIDIX basis set for all atoms, ensuring accurate modeling of molecular orbitals and geometry. To simulate the solvent environment, the polarizable continuum model (PCM) was applied using parameters suitable for DMSO, reflecting realistic experimental conditions. From the optimized molecular structures, key electronic properties such as the energies of the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) were extracted, providing insight into the ligand’s reactivity and stability. 3.3.1. Density Functional Theory Results The energy gap (ΔE), defined as the difference between the Highest Occupied Molecular Orbital (HOMO) and the Lowest Unoccupied Molecular Orbital (LUMO), is a vital parameter for assessing the stability and reactivity of a molecule. In this study, DFT calculations were carried out to analyze the electronic properties of the synthesized stilbene derivative, revealing a HOMO-LUMO energy gap of 3.948 eV, as illustrated in Fig. 12 and summarized in Table 2 . A large energy gap typically indicates high molecular stability, reflecting the significant energy required for electronic excitation. This corresponds to low chemical reactivity, suggesting that the compound is less prone to nucleophilic or electrophilic attack. Such stability is often correlated with enhanced biological activity, as stable molecules tend to form stronger and more specific binding interactions in biological environments. The HOMO is predominantly localized over specific regions of the molecule, highlighting potential sites susceptible to oxidation, whereas the LUMO exhibits delocalization that points to possible electron-accepting sites. This electronic configuration may play a crucial role in the compound’s antifungal activity by influencing its interactions with fungal proteins. Table 2: HOMO-LUMO Surface Structures energy gap (ΔE = 3.948 eV) of the stilbene derivative, highlighting its electronic stability and potential reactivity. Compound Ids E LUMO (eV) E HOMO (eV) ΔE (gap) = E LUMO – E HOMO (eV) 1 -0.997 -4.945 3.948 Overall, the DFT results confirm that the stilbene derivative exhibits high electronic stability, which may enhance its effectiveness as a lead molecule for antifungal drug development. 3.4. Biological Evaluation Of 2, 2', 4, 4', 5, 5'-Hexahydroxy Stilbene Screening for the antifungal target of the selected ligand involved obtaining the three-dimensional (3D) crystal structure of Candida albicans protein (PDB ID: 4HOE) from the Protein Data Bank. The structure was prepared using the Protein Preparation Wizard in the Schrödinger Suite (Maestro v.2023). During preparation, bond orders were assigned, the hydrogen-bonding network was optimized, and missing hydrogen atoms were added to the structure. Water molecules located more than 5 Å away from the binding pocket were removed to avoid interference, while metal ions critical to maintaining the enzyme’s structural integrity were retained. The protein structure was then energy-minimized using the OPLS4 force field, ensuring proper geometry and stabilization of the enzyme. The active site for molecular docking was defined around the heme-binding region, preserving the native ligand and key catalytic residues to facilitate accurate ligand-receptor interaction studies. 3.4.1. Antifungal Activity The antifungal potential of the synthesized compounds was evaluated against Candida albicans , a well-known human pathogenic fungus. The fungal strains were sourced from a certified Microbial Type Culture Collection and Gene Bank (MTCC). Antifungal activity was assessed using the agar dilution method, with compound concentrations ranging from 50 to 200 µg/mL. The minimal inhibitory concentration (MIC) of each test compound was determined following established protocols [10]. Itraconazole, at a dose of 5 grams, was used as a standard reference antifungal agent for comparison. MIC values were specifically determined for yeast isolates that demonstrated sensitivity in the initial disc diffusion assay. The inoculum was prepared from 12-hour-old broth cultures of Candida albicans . Test compounds were initially dissolved in ethanol and diluted to a maximum concentration of 200 µg/mL. Serial two-fold dilutions were then prepared to achieve concentrations of 50, 100, 150, and 200 µg/mL. The MIC determination was carried out by applying these dilutions onto Potato Dextrose Agar (PDA) plates, designated as P1, followed by incubation and evaluation of fungal growth inhibition. 3.4.2. Antifungal Activity by Disc Diffusion Method The disc diffusion assay of 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene against Candida albicans demonstrated a concentration-dependent antifungal effect, with zones of inhibition ranging from 0.8 mm at 200 µg/mL to 18.2 mm at 1000 µg/mL. For comparison, Itraconazole (10 µg/mL) was used as a standard antifungal agent, exhibiting a zone of inhibition of 16.1 mm. These results, illustrated in Fig. 13 and detailed in Table 3, highlight the promising antifungal activity of the synthesized compound, surpassing the efficacy of the standard drug at higher concentrations. Table 3 Minimum inhibition concentration values Compound Concentration Candida albicans 2, 2', 4, 4', 5, 5'-Hexahydroxystilbene 200 µg/ml 0.8 mm 400 µg/ml 2.4 mm 800 µg/ml 15.1mm 1000 µg/ml 18.2 mm Itraconazole 10 µg/ml 16.1mm A comparison study [22] on benzo[4, 5]imidazo[1,2-d][1, 2, 4]triazine derivatives demonstrated significant antifungal activity, primarily attributed to their inhibition of ergosterol biosynthesis, which disrupts fungal cell membrane integrity. In contrast, 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene exhibits strong antifungal properties through a distinct mechanism. The multiple hydroxyl groups present in Hexahydroxy stilbene play a crucial role in generating reactive oxygen species (ROS), which induce oxidative stress and trigger apoptosis in fungal cells, thereby contributing to the compound’s antifungal efficacy [23]. Besides ROS generation, these hydroxyl groups also interact with the fungal membrane, causing destabilization of the lipid bilayers. This interaction leads to ion leakage and membrane disruption, key factors enhancing its antifungal activity [24]. The dual-action mechanism, involving both oxidative stress and membrane destabilization, is central to the compound’s effectiveness against fungal pathogens. Furthermore, the structure of 2,2',4,4',5,5'-Hexahydroxy stilbene shares similarities with other antifungal agents such as benzo[4, 5]imidazo[1,2-d][1, 2, 4]triazine derivatives [25]. Both compound classes disrupt fungal membranes and inhibit critical biosynthetic pathways. However, the hexahydroxylation of stilbene enhances its hydrophilicity, which improves diffusion into fungal cells, increasing solubility, bioavailability, and ultimately, its antifungal potency. 4. Conclusion This study highlights the strong antifungal potential of the synthetic compound 2,2′,4,4′,5,5′-Hexahydroxy stilbene, highlighting its promise as a novel therapeutic agent. Using an integrated approach—combining chemical synthesis, spectral characterization, molecular modeling, and biological evaluation—the research confirms the significance of polyphenolic structures in antifungal activity. Analytical techniques (FTIR, NMR, XRD, and mass spectrometry) validated the compound’s structure and purity. Computational docking showed strong binding to fungal protein targets, outperforming standard ligands, while DFT calculations indicated favorable electronic stability. In vitro assays demonstrated potent antifungal activity, comparable or superior to Itraconazole. Its dual mechanism—ROS generation and membrane disruption—offers a distinct advantage over conventional drugs. These findings support further investigation of hydroxy stilbene derivatives as promising antifungal candidates amid rising drug resistance. Declarations Acknowledgement We take opportunity to thank the University of Mysore and Maulana Azad National Urdu University for giving an opportunity to execute the research. Data availability All data generated or analyzed during this study are included in this published article [and its Supplementary Information file]. Credit Authorship Contribution Statement Gopal Krishna Murthy : Writing – review & editing, Writing – original draft, Methodology, Investigation, Formal analysis. Revanasidappa : Writing – review & editing, Writing – original draft, Visualization, Validation, Methodology, Investigation, Formal analysis. Shivaprakash.S : Writing – review & editing, Writing – original draft, Methodology, Investigation. Vennila Kailasam Natesan : Writing – review & editing, Writing – original draft, Formal analysis, Data curation. Mohammed Saleh Al Ansari : Writing – review & editing. Suhail Ahmad : Writing – review & editing, Writing – original draft, Methodology, Investigation, Formal analysis. Disclosure Statement The authors declare that there are no conflicts of interest regarding the publication of this paper. Funding No funding was received for the preparation of this article. References S. Sepehri, M. Khedmati, F. Yousef-Nejad, M. Mahdavi, Medicinal chemistry perspective on the structure–activity relationship of stilbene derivatives, RSC Adv. D4RA02867H (2024). https://doi.org/10.1039/D4RA02867H D. Šunjka, Š. Mechora, An alternative source of biopesticides and improvement in their formulation—recent advances, Plants (Basel) 11 (22) (2022) 3172. https://doi.org/10.3390/plants11223172 E.W.C. Chan, C.W. Wong, Y.H. Tan, J.P.Y. Foo, S.K. Wong, H.T. Chan, Resveratrol and pterostilbene: A comparative overview of their chemistry, biosynthesis, plant sources, and pharmacological properties, J. Appl. Pharm. Sci. 9 (7) (2019) 17–23. https://doi.org/10.7324/JAPS.2019.90717 F. Caruso, L. Mendoza, P. Castro, M. Cotoras, M. Aguirre, B. Matsuhiro, M. Isaacs, M. Rossi, A. Viglianti, Antifungal activity of resveratrol against Botrytis cinerea is improved using 2-furyl derivatives, PLoS One 6 (10) (2011) e25421. https://doi.org/10.1371/journal.pone.0025421 L. Mattio, G. Catinella, M. Iriti, L. Vallone, Inhibitory activity of stilbenes against filamentous fungi, Plants (Basel) 10 (4) (2021) 773. https://doi.org/10.3390/plants10040773 A. Hammerbacher, S.G. Ralph, J. Bohlmann, T.M. Fenning, J. Gershenzon, A. Schmidt, Biosynthesis of the major tetrahydroxystilbenes in spruce, astringin and isorhapontin, proceeds via resveratrol and is enhanced by fungal infection, Plant Physiol. 157 (3) (2011) 876–890. https://doi.org/10.1104/pp.111.179325 N. Patel, V. Karkhanis, P. Patel, Synthesis and biological evaluation of some piperazine derivatives as anti-inflammatory agents, J. Drug Deliv. Ther. 9 (4-s) (2019) 353–358. https://doi.org/10.22270/jddt.v9i4-s.3327 C.F.G.C. Geraldes, Introduction to infrared and Raman-based biomedical molecular imaging and comparison with other modalities, Molecules 25 (23) (2020) 5547. https://doi.org/10.3390/molecules25235547 V. Andrei, A.G.N. Wetie, I. Mihai, C.C. Andrei, Detection of biomedically relevant stilbenes from wines by mass spectrometry, in: J. Bousquet, M. Fountoulakis (Eds.), Advancements of Mass Spectrometry in Biomedical Research, Springer, Cham, 2014, pp. 361–382. https://doi.org/10.1007/978-3-319-06068-2_17 R. Gunawan, A. Nandiyanto, I. Ijost, How to read and interpret ¹H-NMR and ¹³C-NMR spectrums, Indones. J. Sci. Technol. 6 (2) (2021) 267–298. https://doi.org/10.17509/ijost.v6i2.34189 J. O, J. Young, True melting point determination, J. Chem. Educ. 18 (2013) 203–208. https://doi.org/10.1333/s00897132500a S. Shivaprakash, G.C. Reddy, J.P. Jasinski, S.P. Millikan, C.E. Duff, C. Glidewell, A sterically congested cis-stilbene and its phosphonium salt precursor, Acta Crystallogr. C Struct. Chem. 71 (6) (2015) 479–484. https://doi.org/10.1107/S2053229615009110 S.K. Burley, C. Bhikadiya, C. Bi, et al., RCSB Protein Data Bank (RCSB.org): delivery of experimentally-determined PDB structures alongside one million computed structure models of proteins from artificial intelligence/machine learning, Nucleic Acids Res. 51 (D1) (2023) D488–D508. https://doi.org/10.1093/nar/gkac1077 T. Egawa, S.-R. Yeh, D. Rousseau, Redox-controlled proton gating in bovine cytochrome c oxidase, PLoS One 8 (2013) e63669. https://doi.org/10.1371/journal.pone.0063669 W. Tian, C. Chen, X. Lei, J. Zhao, Y. Li, Q. Wang, CASTp 3.0: Computed atlas of surface topography of proteins, Nucleic Acids Res. 46 (W1) (2018) W363–W367. https://doi.org/10.1093/nar/gky473 C. Colovos, T.O. Yeates, Verification of protein structures: patterns of nonbonded atomic interactions, Protein Sci. 2 (9) (1993) 1511–1519. https://doi.org/10.1002/pro.5560020916 O. Trott, A.J. Olson, AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading, J. Comput. Chem. 31 (2) (2010) 455–461. https://doi.org/10.1002/jcc.21334 A.C. Wallace, R.A. Laskowski, J.M. Thornton, LIGPLOT: A program to generate schematic diagrams of protein-ligand interactions, Protein Eng. 8 (2) (1995) 127–134. https://doi.org/10.1093/protein/8.2.127 S. Forli, R. Huey, M.E. Pique, M.F. Sanner, D.S. Goodsell, A.J. Olson, Computational protein–ligand docking and virtual drug screening with the AutoDock suite, Nat. Protoc. 11 (5) (2016) 905–919. https://doi.org/10.1038/nprot.2016.051 G. Li, X. Ma, L. Deng, X. Zhao, Y. Wei, Z. Gao, J. Jia, J. Xu, C. Sun, Fresh garlic extract enhances the antimicrobial activities of antibiotics on resistant strains in vitro, J. Infect. Microbiol. 10 (2025) 14814. https://doi.org/10.5812/jjm.14814 A.V. Snezhkina, A.V. Kudryavtseva, O.L. Kardymon, M.V. Savvateeva, N.V. Melnikova, G.S. Krasnov, A.A. Dmitriev, ROS generation and antioxidant defense systems in normal and malignant cells, Oxid. Med. Cell. Longev. 2019 (2019) 6175804. https://doi.org/10.1155/2019/6175804 P. Nakhaei, R. Margiana, D.O. Bokov, W.K. Abdelbasset, M.A. Jadidi Kouhbanani, R.S. Varma, F. Marofi, M. Jarahian, N. Beheshtkhoo, Liposomes: structure, biomedical applications, and stability parameters with emphasis on cholesterol, Front. Bioeng. Biotechnol. 9 (2021) 705886. https://doi.org/10.3389/fbioe.2021.705886 T. Kon, N. Nemoto, T. Oshima, A. Yamagishi, Effects of a squalene epoxidase inhibitor, terbinafine, on ether lipid biosyntheses in a thermoacidophilic archaeon, Thermoplasma acidophilum, J. Bacteriol. 184 (5) (2002) 1395–1401. https://doi.org/10.1128/JB.184.5.1395-1401.2002 M. Krysa, M. Szymańska-Chargot, A. Zdunek, FT-IR and FT-Raman fingerprints of flavonoids – a review, Food Chem. 393 (2022) 133430. https://doi.org/10.1016/j.foodchem.2022.133430 K. Jomova, R. Raptova, S.Y. Alomar, S.H. Alwasel, E. Nepovimova, K. Kuca, M. Valko, Reactive oxygen species, toxicity, oxidative stress, and antioxidants: chronic diseases and aging, Arch. Toxicol. 97 (10) (2023) 2499–2574. https://doi.org/10.1007/s00204-023-03562-9 Additional Declarations No competing interests reported. Supplementary Files SupplementaryDataInformation.docx floatimage1.jpeg Graphical Abstract 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-6755724","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":472102422,"identity":"c12b3b49-28a7-479f-a12c-766712a94e22","order_by":0,"name":"Gopal Krishna Murthy H.R.","email":"","orcid":"","institution":"University of Mysore","correspondingAuthor":false,"prefix":"","firstName":"Gopal","middleName":"Krishna Murthy","lastName":"H.R.","suffix":""},{"id":472102423,"identity":"2052efc3-7e88-4ac6-aec1-67edd0f13c1a","order_by":1,"name":"Revanasiddappa H.D.","email":"","orcid":"","institution":"University of Mysore","correspondingAuthor":false,"prefix":"","firstName":"Revanasiddappa","middleName":"","lastName":"H.D.","suffix":""},{"id":472102426,"identity":"0def6254-3a45-4151-8837-2713cee922a2","order_by":2,"name":"Shivaprakash S","email":"","orcid":"","institution":"Vittal Mallya Scientific Research Foundation","correspondingAuthor":false,"prefix":"","firstName":"Shivaprakash","middleName":"","lastName":"S","suffix":""},{"id":472102427,"identity":"f4c82f29-8a10-4aca-8869-cef4938b789f","order_by":3,"name":"Vennila Kailasam Natesan","email":"","orcid":"","institution":"PSNA College of Engineering and Technology","correspondingAuthor":false,"prefix":"","firstName":"Vennila","middleName":"Kailasam","lastName":"Natesan","suffix":""},{"id":472102428,"identity":"4a91fb93-efbd-4ff4-98e6-952f3a01c33b","order_by":4,"name":"Mohammed Saleh Al Ansari Al Ansari","email":"","orcid":"","institution":"University of Bahrain","correspondingAuthor":false,"prefix":"","firstName":"Mohammed","middleName":"Saleh Al Ansari Al","lastName":"Ansari","suffix":""},{"id":472102429,"identity":"03f2a4e2-87e6-42d6-b98d-0fc39afd2847","order_by":5,"name":"Suhail Ahmad","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABDklEQVRIie2PsUrEQBCGNxxstZh2CzGvMHCwWITcg9jMsXA2ObEUPHCtrhFs40Nca71hQZvDtJFrcr1IrrtKnGxhl8RScL9iBob5+GcYCwT+JIIxvGannMWNPXylNInu7bgCTHAmorLgi04x4wqDroqJE9z52aCSFMuybSATJ/HGloWoss3aUcoqvehToL7SEkELLj/RtnKnn7dzUl4WS9OnyBxImZBioSxgp5UlJTKuV0mKfHpEuPOKE/imVbUfVlidK0qhx+MHqtZmqh5Jge2HOkd4pRSO5ZPRqGpKwYFfknU+fT/e3J4lj861B5PNVHW5b9pV2n/YDxJ9m/tNHF3viK1vs18tBwKBwL/iG02iZXdpvRsiAAAAAElFTkSuQmCC","orcid":"","institution":"Maulana Azad National Urdu University","correspondingAuthor":true,"prefix":"","firstName":"Suhail","middleName":"","lastName":"Ahmad","suffix":""}],"badges":[],"createdAt":"2025-05-27 05:53:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6755724/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6755724/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":84819533,"identity":"591b0b98-5d09-4ae7-ace9-ba0626eb1cd3","added_by":"auto","created_at":"2025-06-17 15:59:58","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":57623,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003e(Z)\u003c/em\u003e-2, 2', 4, 4', 5, 5'- Hexahydroxy stilbene\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/5921975b94c67195107f5b83.png"},{"id":84819537,"identity":"27f2db06-ba2e-4af3-8368-4113a300a89e","added_by":"auto","created_at":"2025-06-17 15:59:58","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":165309,"visible":true,"origin":"","legend":"\u003cp\u003eSynthesis of (Z)-2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/0ba79caf3a3f8c9b0e4b1d12.png"},{"id":84819536,"identity":"f8b54073-4af3-441e-b20b-a80a450fd761","added_by":"auto","created_at":"2025-06-17 15:59:58","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":419029,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR analysis of (Z) - 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/efc82841ecd97424bf79041c.png"},{"id":84819542,"identity":"319fa874-6acc-4d85-8b4c-b3b1d7b264fd","added_by":"auto","created_at":"2025-06-17 15:59:58","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":207122,"visible":true,"origin":"","legend":"\u003cp\u003eMass spectrometry analysis of 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/68606db4066bca46f07deff8.png"},{"id":84821866,"identity":"b2eabf0d-e6e5-4a22-a183-df3a13ab2942","added_by":"auto","created_at":"2025-06-17 16:15:58","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":268118,"visible":true,"origin":"","legend":"\u003cp\u003e\u003csup\u003e1\u003c/sup\u003eH-NMR analysis of 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/3a8030a60b1b4dc0cc0994c7.png"},{"id":84819550,"identity":"2aff4512-8ccb-47af-bbca-e96c657e97fc","added_by":"auto","created_at":"2025-06-17 15:59:58","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":389153,"visible":true,"origin":"","legend":"\u003cp\u003e\u003csup\u003e1\u003c/sup\u003eH-NMR analysis of 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/57f2318c68c3dc7a53807dc8.png"},{"id":84820342,"identity":"aae55757-1312-45b2-b71a-3377e86ff375","added_by":"auto","created_at":"2025-06-17 16:07:58","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":219555,"visible":true,"origin":"","legend":"\u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC-NMR analysis of 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/bb725deb6283384a2c3ca2e8.png"},{"id":84820343,"identity":"7c5fe81a-0d2c-43fc-ace9-b73fc5ed1471","added_by":"auto","created_at":"2025-06-17 16:07:58","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":166428,"visible":true,"origin":"","legend":"\u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC-NMR analysis of 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/7796809183e9d44a5f7bcad9.png"},{"id":84821868,"identity":"dfdf4763-b9cf-4803-89b9-e1b07041d60e","added_by":"auto","created_at":"2025-06-17 16:15:59","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":88737,"visible":true,"origin":"","legend":"\u003cp\u003eXRD analysis of 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/e0d3de590fa117f98ff50cfb.png"},{"id":84820339,"identity":"6c55bcad-f1af-48c0-9317-8c17d6fb6836","added_by":"auto","created_at":"2025-06-17 16:07:58","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":876652,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe Ramachandran plot generated from RAMPAGE. The Ramachandran plot representing energetically allowable regions for backbone dihedral angles ψ vs φ amino acid residues in selected protein structure (A) \u003c/strong\u003eThe structure\u003cstrong\u003e \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eCandida albicans\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e (PDB ID:4HOE)\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/b7c37f3cc4a034f8d32e898b.png"},{"id":84819579,"identity":"30dfdc86-7110-4ca7-92c8-299a0942445f","added_by":"auto","created_at":"2025-06-17 15:59:59","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":593636,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e2-dimensional visualisation of molecular docking of Candida albicans (PDB ID: 4HOE) \u003c/strong\u003ehighlighting hydrogen bonds, hydrophobic interactions where (A) and (B) The molecular docking interactions of hydroxy stilbene with the fungal protein (PDB ID: 4HOE)\u003c/p\u003e","description":"","filename":"11.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/6d1c57cec717f0dee4a43a4d.png"},{"id":84819568,"identity":"5b1e2524-5ad7-479d-a0a6-c13b7ab54892","added_by":"auto","created_at":"2025-06-17 15:59:59","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":603168,"visible":true,"origin":"","legend":"\u003cp\u003eHOMO-LUMO Surface Structuresenergy gap (ΔE = 3.948 eV) of the stilbene derivative, highlighting its electronic stability and potential reactivity.\u003c/p\u003e","description":"","filename":"12.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/242af65f8b99d8ec287d7589.png"},{"id":84819564,"identity":"69330553-42ed-49f2-9af7-24418f770dbe","added_by":"auto","created_at":"2025-06-17 15:59:58","extension":"png","order_by":13,"title":"Figure 13","display":"","copyAsset":false,"role":"figure","size":289232,"visible":true,"origin":"","legend":"\u003cp\u003eThe inhibition of \u003cem\u003eCandida albicans\u003c/em\u003e treated with 2, 2', 4, 4', 5, 5'-Hexahydroxystilbene at varying concentrations (200, 400, 800, 1000 µg/ml) and Itraconazole (10 µg/ml) as a positive control.\u003c/p\u003e","description":"","filename":"13.png","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/4f378eac5195c16685f5c19b.png"},{"id":85515436,"identity":"9baa9b84-4059-4ee5-a1cf-8e80e105f832","added_by":"auto","created_at":"2025-06-26 17:46:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":6578341,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/24499ef9-5726-4582-9512-3892a8ba0e49.pdf"},{"id":84819539,"identity":"d4ac3a38-57d6-4782-9f8d-d203f48c7e80","added_by":"auto","created_at":"2025-06-17 15:59:58","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1394891,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryDataInformation.docx","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/854a12ecec6cdbb011f17270.docx"},{"id":84820338,"identity":"e1414bab-4723-430c-aaea-f502befc780b","added_by":"auto","created_at":"2025-06-17 16:07:58","extension":"jpeg","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":381488,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphical Abstract\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-6755724/v1/0bfc2e84e9dd7d536d9256cb.jpeg"}],"financialInterests":"No competing interests reported.","formattedTitle":"Design, Synthesis, Structural, and Computational Studies of (Z)-2,2′,4,4′,5,5′-Hexahydroxy Stilbene as a Potential Antifungal Agent","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eStilbenes are a distinct class of phenylpropanoid compounds characterized by a 1,2-diphenylethylene backbone. Despite being a relatively small subclass within the broad phenylpropanoid family, they are widely distributed in various plant species and serve as essential components in plant defense mechanisms [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Most stilbene derivatives, collectively referred to as stilbenoids, are naturally synthesized by plants under stress conditions and act as phytoalexins\u0026mdash;defensive compounds produced in response to pathogenic attacks. Beyond their role in plant immunity, stilbenoids have garnered increasing attention for their diverse biological activities, particularly their ability to interfere with microbial systems. Studies have shown that these compounds can effectively disrupt microbial biofilms and attenuate key virulence factors in bacteria and fungi [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. These properties make stilbenoids promising candidates for the development of alternative antimicrobial agents, especially in the face of the rising global challenge of drug resistance. Moreover, the environmentally benign nature of these plant-based compounds has led to their growing application in sustainable agriculture, where they offer a safer alternative to synthetic pesticides, combining efficacy with ecological safety [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAmong naturally occurring stilbenoids, \u003cem\u003etrans\u003c/em\u003e-resveratrol is one of the most well-studied and biologically active members. Found abundantly in grapes, berries, and peanuts, \u003cem\u003etrans\u003c/em\u003e-resveratrol has been shown to possess significant antifungal properties [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. One of the most notable targets of resveratrol is \u003cem\u003eBotrytis cinerea\u003c/em\u003e, a fungal pathogen responsible for gray mold in fruits, leading to considerable post-harvest losses in agriculture. The antifungal mechanism of \u003cem\u003etrans\u003c/em\u003e-resveratrol is believed to involve disruption of fungal cell membranes, inhibition of metabolic enzymes, and interference with cellular respiration [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. These effects result in impaired fungal growth and survival, highlighting the potential of stilbenoids as biocontrol agents. In addition to their direct antimicrobial action, resveratrol and related stilbenoids may exert synergistic effects when used alongside conventional antifungal agents, potentially reducing the required dosage and limiting resistance development.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eStructurally, stilbenes can exist in two stereoisomeric forms\u0026mdash;\u003cem\u003eE\u003c/em\u003e- (trans) and \u003cem\u003eZ\u003c/em\u003e- (cis)\u0026mdash;which exhibit different physicochemical and biological properties. The ability of stilbenes to undergo \u003cem\u003eE/Z\u003c/em\u003e isomerization adds an extra layer of complexity to their behavior, as this transformation can significantly alter their molecular geometry and reduce their biological efficacy [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. While \u003cem\u003etrans\u003c/em\u003e-stilbenes are thermodynamically more stable and have been extensively investigated, \u003cem\u003ecis\u003c/em\u003e-stilbenes have attracted increasing interest in recent years due to their distinct structural features. In the \u003cem\u003ecis\u003c/em\u003e-configuration, the two phenyl rings are positioned on the same side of the ethylene double bond, giving rise to a unique three-dimensional geometry that affects both reactivity and interaction with biological targets [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. These characteristics make \u003cem\u003ecis\u003c/em\u003e-stilbenes particularly valuable in photochemical studies, materials science, and medicinal chemistry. Importantly, structural modifications\u0026mdash;such as the introduction of hydroxyl groups\u0026mdash;have been shown to enhance the biological activity of stilbenes. Hydroxylation not only improves the compound\u0026rsquo;s hydrophobic interactions with lipid membranes but also increases its capacity to induce oxidative stress and inhibit fungal enzymatic systems [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn light of these promising attributes, the present study focused on the synthesis of a novel hydroxylated \u003cem\u003ecis\u003c/em\u003e-stilbene compound, (Z)-2,2\u0026prime;,4,4\u0026prime;,5,5\u0026prime;-hexahydroxy stilbene, achieved through a one-step demethylation reaction from the corresponding methoxylated precursor. This synthetic route provides an efficient strategy to obtain the desired hydroxylated derivative while retaining the \u003cem\u003eZ\u003c/em\u003e-configuration. The newly synthesized compound was comprehensively characterized using FTIR spectroscopy, \u0026sup1;H and \u0026sup1;\u0026sup3;C nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry (MS), and single-crystal X-ray diffraction, which confirmed its molecular geometry and stereochemistry. To explore the electronic and molecular properties of the compound, density functional theory (DFT) calculations were performed, offering valuable insights into its stability and reactivity. Furthermore, molecular docking studies were conducted to investigate the interaction of the compound with fungal target proteins, providing evidence of strong binding affinity and suggesting possible modes of antifungal action. These findings not only support the structural and functional integrity of the synthesized compound but also underscore its potential as a promising lead molecule for the development of new antifungal therapeutics.\u003c/p\u003e"},{"header":"2. Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. General methods\u003c/h2\u003e \u003cp\u003eAll chemicals used in this study were of Laboratory Reagent (LR) grade and were procured from commercial suppliers. Solvents employed were of chemical grade and used without further purification unless specified otherwise. Thin-layer chromatography (TLC) was conducted using aluminium sheets pre-coated with silica gel 60 F254 (Merck), and the visualization of spots was performed under UV light at 254 nm. Reaction progress and compound purity were routinely monitored using TLC.\u003c/p\u003e \u003cp\u003eAnalytical High-Performance Liquid Chromatography (HPLC) was carried out using a Shimadzu CLASS-VP system comprising LC-10AT VP high-pressure binary pumps, an SPD-M10A VP photodiode array detector, a CTO-10AS VP column oven, and an SCL-10A VP system controller. The separation of analytes was achieved on a reversed-phase Atlantis-T3 column (5.0 \u0026micro;m, 4.6 \u0026times; 150 mm) under isocratic conditions using a mobile phase of acetonitrile and water (50:50, v/v). The elution was monitored at 290 nm with a UV detector. Chromatographic purification of the synthesized compounds was performed by open-column chromatography using Merck silica gel (Grade 7734, 70\u0026ndash;230 mesh). Elution was carried out using ethyl acetate and hexane mixtures in varying proportions, and fractions were combined based on TLC analysis.\u003c/p\u003e \u003cp\u003eMelting points of the synthesized compounds were determined using an Arco melting point apparatus fitted with a calibrated thermometer, with a measurement accuracy of \u0026plusmn;\u0026thinsp;1\u0026deg;C. Samples were placed in sealed capillary tubes and heated gradually, and the values reported are uncorrected.\u003c/p\u003e \u003cp\u003eFourier-transform infrared (FTIR) spectra were recorded on a PerkinElmer Spectrum IR instrument (version 10.7.2) over a spectral range of 4000\u0026ndash;500 cm⁻\u0026sup1;. The samples were finely ground and pelletized with potassium bromide (KBr) before analysis. Key absorption bands were examined to identify functional groups and verify structural features.\u003c/p\u003e \u003cp\u003eMass spectrometry was performed using electrospray ionization in positive ion mode (ESI+). For each analysis, approximately 1 mg of the sample was dissolved in methanol and injected into the mass spectrometer. Molecular ion peaks and fragmentation patterns were used to confirm molecular weights and assess structural consistency with the proposed structures.\u003c/p\u003e \u003cp\u003e\u0026sup1;H and \u0026sup1;\u0026sup3;C nuclear magnetic resonance (NMR) spectra were recorded on a Bruker spectrometer operating at 400 MHz and 100 MHz, respectively. Deuterated chloroform (CDCl₃) served as the solvent, with tetramethylsilane (TMS) used as the internal standard. Chemical shifts were expressed in parts per million (δ), and detailed analyses of multiplicities, coupling constants, and integration values were performed to elucidate the molecular structure and confirm compound purity.\u003c/p\u003e \u003cp\u003eX-ray diffraction (XRD) patterns were obtained using a Rigaku diffractometer employing Cu Kα radiation (λ\u0026thinsp;=\u0026thinsp;1.5406 \u0026Aring;). Data were collected across a 2θ range of 5\u0026deg; to 80\u0026deg; at a scanning rate of 1\u0026deg; per minute. The diffraction data were analyzed to assess crystallinity and identify the phases present in the synthesized materials by comparison with standard patterns.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Synthesis of (Z) \u0026minus;\u0026thinsp;2, 2', 4, 4', 5, 5'- Hexahydroxy stilbene\u003c/h2\u003e \u003cp\u003eThe synthetic procedure was carried out according to the scheme illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. To a stirred solution of triethylamine (TEA) (16.86 g, 166.67 mmol) in chlorobenzene (7.5 g, 66.6 mmol) under a nitrogen atmosphere at 0\u0026ndash;5\u0026deg;C, anhydrous aluminium chloride (AlCl₃) (13.32 g, 99.9 mmol) was added slowly in small portions over 30\u0026ndash;40 minutes. After complete addition, the reaction mixture was allowed to reach ambient temperature and stirred for 30\u0026ndash;32 minutes. The temperature was then gradually raised to 60\u0026deg;C and maintained at this level for 1 hour with continuous stirring. Subsequently, a solution of (E)-2,2\u0026prime;,4,4\u0026prime;,5,5\u0026prime;-hexamethoxystilbene (2.0 g, 5.55 mmol) in chlorobenzene (7.5 g, 66.6 mmol) was added dropwise over 30 minutes while maintaining the temperature at 60\u0026ndash;65\u0026deg;C, followed by an additional 30 minutes of stirring. The progress of the reaction was monitored by thin-layer chromatography (TLC) using silica gel plates pre-coated with silica gel 60 F254 (Merck) and visualized under UV light at 254 nm.\u003c/p\u003e \u003cp\u003eUpon completion, the reaction mixture was cooled to room temperature and quenched by pouring into ice water (150 g). The resulting biphasic mixture was extracted twice with methylene dichloride (MDC) (2 \u0026times; 100 mL). The combined organic layers were washed with water (50 mL) and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to afford a crude residue. This crude mass was subjected to column chromatography over silica gel (220\u0026ndash;400 mesh) using hexane:ethyl acetate (8:2) as the mobile phase to yield the desired product, (Z)-2,2\u0026prime;,4,4\u0026prime;,5,5\u0026prime;-hexahydroxystilbene (0.616 g, 2.22 mmol) in 40.43% yield as an off-white solid. The product exhibited a melting point of 134\u0026ndash;135\u0026deg;C.\u003c/p\u003e \u003cp\u003eSpectroscopic data confirmed the structure and purity of the product. \u003csup\u003e1\u003c/sup\u003eH NMR (CdCl₃, 400 MHz): δ 7.716\u0026ndash;7.662 (multiplet, 5H), 7.589\u0026ndash;7.541 (multiplet, 3H), 7.503\u0026ndash;7.456 (multiplet, 4H). \u003csup\u003e13\u003c/sup\u003eC NMR (CDCl₃, 100 MHz): δ 133.00, 132.15, 132.05, 131.97, 131.95, 128.58, 128.45. Mass spectrometric analysis (ESI+) showed a peak at m/z 279 [M\u0026thinsp;+\u0026thinsp;2], consistent with the calculated molecular ion for C₁₄H₁₂O₆ (277.08) [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results and Discussion","content":"\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Spectral analysis of 2, 2', 4, 4', 5, 5'-hexahydroxy stilbene\u003c/h2\u003e \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e \u003ch2\u003e3.1.1. FTIR analysis\u003c/h2\u003e \u003cp\u003eThe FTIR spectrum of the synthesized (Z)-2,2\u0026prime;,4,4\u0026prime;,5,5\u0026prime;-hexahydroxystilbene, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, exhibits characteristic peaks confirming the presence of key functional groups integral to its structure. A prominent absorption band at 3055.89 cm⁻\u0026sup1; corresponds to the O\u0026ndash;H stretching vibration typical of phenolic hydroxyl groups, indicating the presence of hydroxyl substituents on the aromatic rings of the stilbene framework. A weak absorption at 1979.00 cm⁻\u0026sup1; may be attributed to overtone or combination bands associated with aromatic C\u0026thinsp;=\u0026thinsp;C stretching. The strong peak at 1590.75 cm⁻\u0026sup1; is assigned to aromatic C\u0026thinsp;=\u0026thinsp;C stretching vibrations, confirming the presence of benzene rings within the stilbene derivative. Additional peaks at 1484.56 cm⁻\u0026sup1; and 1437.76 cm⁻\u0026sup1; further validate the aromatic ring vibrations.\u003c/p\u003e \u003cp\u003eThe absorption band at 1395.33 cm⁻\u0026sup1; suggests O\u0026ndash;H in-plane bending, characteristic of phenolic compounds. Aromatic C\u0026ndash;H in-plane bending vibrations are indicated by the peak at 1311.62 cm⁻\u0026sup1;. The peaks observed at 1182.87 cm⁻\u0026sup1; and 1119.83 cm⁻\u0026sup1; correspond to C\u0026ndash;O stretching vibrations, supporting the presence of hydroxyl functionalities. Further absorptions at 1071.08 cm⁻\u0026sup1; and 1025.84 cm⁻\u0026sup1; are attributed to C\u0026ndash;H bending and possibly secondary C\u0026ndash;O stretching modes. The range from 995.08 cm⁻\u0026sup1; to 694.72 cm⁻\u0026sup1; displays bands corresponding to C\u0026ndash;H out-of-plane bending, confirming a substituted aromatic system. Collectively, the FTIR data affirm the presence of hydroxy groups, aromatic rings, and the stilbene core, all of which are essential to the compound\u0026rsquo;s bioactivity.\u003c/p\u003e \u003cp\u003eHydroxy stilbenes, particularly (Z)-2,2\u0026prime;,4,4\u0026prime;,5,5\u0026prime;-hexahydroxystilbene, are well-known for their polyphenolic nature, which significantly contributes to their antifungal properties. These compounds induce reactive oxygen species (ROS) generation, leading to oxidative stress and apoptosis in fungal cells [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Moreover, the phenolic hydroxyl groups interact with lipid bilayers, destabilizing fungal membranes and causing leakage of essential ions [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Stilbene derivatives are also reported to inhibit fungal squalene epoxidase and other enzymes involved in ergosterol biosynthesis, crucial for fungal cell membrane integrity [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn our FTIR analysis, the prominent hydroxyl stretching vibration observed at 3055.89 cm⁻\u0026sup1; and the C\u0026ndash;O stretching bands at 1182.87 cm⁻\u0026sup1; and 1119.83 cm⁻\u0026sup1; correspond directly to the functional groups known to play a crucial role in antifungal activity. These vibrational bands are indicative of phenolic hydroxyl groups and ether linkages, which are integral to the bioactive profile of hydroxy stilbene derivatives. A similar study reported by [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e] observed FTIR peaks in the 3050\u0026ndash;3500 cm⁻\u0026sup1; region for hydroxy stilbene compounds exhibiting antifungal properties. They also highlighted that absorption bands in the 1590\u0026ndash;1400 cm⁻\u0026sup1; range were associated with aromatic and phenolic structures, findings that are consistent with the present analysis.\u003c/p\u003e \u003cp\u003eThe extensive hexahydroxylation of the stilbene core significantly enhances the hydrophilicity of the molecule, facilitating improved diffusion through fungal cell walls and membranes. Additionally, the presence of multiple hydroxyl groups contributes to the compound\u0026rsquo;s antioxidant activity and its ability to disrupt fungal membrane integrity, which underpins its efficacy as an antifungal agent [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e3.1.2. Mass Spectral Analysis\u003c/h2\u003e \u003cp\u003eThe mass spectrometric analysis depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e provides detailed insights into the molecular composition and fragmentation pattern of the synthesized hexahydroxystilbene compound. The observed peaks at m/z 279.57 and 280.59 likely correspond to the [M\u0026ndash;H]⁺ and [M\u0026thinsp;+\u0026thinsp;H]⁺ ion species of the base stilbene structure, respectively. The minor difference between these two peaks can be attributed to isotopic distribution or the presence of minor adducts formed during ionization. Peaks at m/z 320.65 and 321.65 suggest fragment ions resulting from hydroxylation or the loss of small substituents such as water (\u0026ndash;H₂O), which is a common fragmentation pathway for phenolic compounds.\u003c/p\u003e \u003cp\u003eHigher m/z values such as 466.49 and 558.02 indicate larger fragment ions or adducts, potentially arising from combinations of hydroxylated moieties or rearranged stilbene cores during ionization. Additionally, high-molecular-weight ions observed at m/z 990.74, 1334.83, and 1461.92 likely correspond to oligomeric species, such as dimers or trimers of the hexahydroxystilbene molecule. These oligomers may be stabilized by hydrogen bonding or other weak intermolecular interactions in the gas phase. Peaks at m/z 1752.21 and 1998.67 may represent even more complex oligomeric forms or non-covalent interactions with the ionization source, which is frequently observed in electrospray ionization positive mode (ESI+) analyses of polyphenolic compounds. These mass spectral features collectively confirm the molecular identity and the propensity of the compound to form stable oligomeric assemblies under the experimental conditions.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe mass spectrometry analysis provides valuable insights into the molecular structure and potential bioactivity of 2,2\u0026prime;,4,4\u0026prime;,5,5\u0026prime;-hexahydroxystilbene. The molecular weight and fragmentation pattern observed are consistent with structural features known to contribute to antifungal activity, as reported in previous studies on related hydroxy stilbene compounds. Hydroxy stilbenes containing multiple hydroxyl groups are recognized for their ability to generate reactive oxygen species (ROS), which induce oxidative stress and trigger apoptosis in fungal cells [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Additionally, phenolic compounds disrupt fungal membrane integrity through interactions with lipid bilayers, compromising membrane stability [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe detection of oligomeric species at m/z 990.74, 1334.83, and 1461.92 may further enhance the antifungal efficacy by promoting cooperative interactions with fungal cell wall components, potentially increasing the compound\u0026rsquo;s binding affinity and disruptive capacity. A prior study by [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e] analyzing the mass spectra of hydroxy stilbene derivatives identified similar high-molecular-weight oligomers, attributing them to dimerization or polymerization processes that are crucial for bioactivity. The peaks observed in the current study align well with these molecular architectures, suggesting that 2,2\u0026prime;,4,4\u0026prime;,5,5\u0026prime;-hexahydroxystilbene possesses structural characteristics favorable for antifungal applications and could serve as a promising bioactive agent.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003e3.1.3. NMR Spectral Analysis\u003c/h2\u003e \u003cp\u003eFigures \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e and \u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e display the ^1H-NMR spectrum of the synthesized compound, revealing characteristic peaks in the aromatic region that confirm the presence of hydroxyl-substituted stilbene moieties. The aromatic protons resonate between 7.45 and 7.71 ppm, consistent with the expected chemical environment of the stilbene aromatic rings. Additionally, a deshielded singlet observed at 4.00 ppm is attributed to hydroxyl protons engaged in hydrogen bonding, which is typical for phenolic hydroxyl groups. This spectral pattern aligns well with previously reported data for hexahydroxystilbene derivatives, further validating the successful synthesis and structural integrity of the compound [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFurther analysis of the \u003csup\u003e13\u003c/sup\u003eC-NMR spectra shown in Figs.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e and \u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e reveals characteristic peaks within the range of 120\u0026ndash;135 ppm, which are indicative of sp\u0026sup2;-hybridized carbons forming the stilbene backbone. Notably, the signals at 128.45 ppm and 132.05 ppm correspond specifically to carbons of the substituted benzene rings. The absence of any aliphatic carbon resonances confirms the high purity of the synthesized compound, ruling out contamination or incomplete reactions. These spectral features are consistent with literature reports on similar polyphenolic stilbene derivatives, supporting the structural assignment of the hexahydroxystilbene [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003e3.1.4. XRD Analysis\u003c/h2\u003e \u003cp\u003eThe X-ray diffraction (XRD) analysis of the synthesized (Z)-2,2\u0026prime;,4,4\u0026prime;,5,5\u0026prime;-hexahydroxystilbene, as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e9\u003c/span\u003e, reveals distinct crystallographic features indicative of its solid-state structure. The diffraction pattern exhibits prominent peaks at specific 2θ angles, reflecting the crystalline nature of the compound. The most intense peak is observed at 23.2512\u0026deg; (2θ) with an intensity of 1,898,414 counts per second (cps), corresponding to the primary crystal plane and demonstrating a high degree of crystallinity. Additional significant peaks at 8.131\u0026deg; and 11.003\u0026deg; (2θ) suggest secondary lattice reflections related to molecular packing arrangements within the crystal lattice. Furthermore, several smaller peaks detected between 15\u0026deg; and 30\u0026deg; (2θ) indicate the presence of multiple crystalline phases or orientations, which is typical of polycrystalline samples. These XRD results collectively confirm the well-defined crystalline nature and structural complexity of the synthesized hexahydroxystilbene derivative.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe full-width at half-maximum (FWHM) values of the diffraction peaks, such as 0.0618\u0026deg; for the main peak at 23.2512\u0026deg; (2θ), indicate narrow peak widths that further confirm the presence of a well-ordered crystal lattice in the synthesized (Z)-2,2\u0026prime;,4,4\u0026prime;,5,5\u0026prime;-hexahydroxystilbene. Minor asymmetry observed in the peaks, exemplified by an asymmetry factor of 1.12 for the primary peak, may be attributed to slight distortions or defects within the crystal lattice. The d-spacing value calculated for the primary peak, approximately 3.822 \u0026Aring;, when compared with standard crystallographic databases, confirms the structural integrity of the compound. These results align well with the expected crystalline nature of the synthesized molecule, corroborating its purity and successful synthesis.\u003c/p\u003e \u003cp\u003eWhen compared with a reference study analyzing the crystallinity of hydroxy stilbenes [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e], the current findings show strong agreement, especially regarding the intense diffraction peak near 23\u0026deg; (2θ). Additional peaks at lower (8.131\u0026deg;) and higher (11.003\u0026deg;) angles observed in our data suggest subtle structural modifications, possibly due to hydroxyl substitutions at specific positions on the stilbene backbone, which influence the crystal packing. The high intensity (1,898,414 cps) and sharpness of the primary peak underscore a high degree of crystallinity, indicative of a pure compound. In contrast, broad or low-intensity peaks would imply amorphous impurities or incomplete crystallization. These observations are consistent with previous findings that emphasize the role of hydroxyl groups in enhancing crystallinity via hydrogen bonding networks.\u003c/p\u003e \u003cp\u003eThe d-spacing values derived from the XRD analysis suggest a tightly packed crystal lattice stabilized by extensive hydrogen bonding interactions among the hydroxyl groups. This network contributes significantly to the rigidity and stability of the crystal structure. The secondary peaks at 8.131\u0026deg; and 11.003\u0026deg; (2θ) likely correspond to layered molecular arrangements within the crystal lattice. The structural integrity and high crystallinity of 2,2\u0026prime;,4,4\u0026prime;,5,5\u0026prime;-hexahydroxystilbene are not only essential for confirming its purity but also play a critical role in its biological activity. According to the referenced study, a direct correlation exists between crystallinity and enhanced bioactivity, as a well-ordered lattice facilitates more effective interaction with biological targets.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Molecular Docking\u003c/h2\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003e3.2.1. Ligand Preparation\u003c/h2\u003e \u003cp\u003eThe synthetic stilbene derivative was initially designed using ChemDraw and subsequently converted into its three-dimensional (3D) structure for computational analysis. Ligand preparation was carried out using LigPrep in the Schr\u0026ouml;dinger suite, where the molecule underwent geometry optimization with the OPLS4 force field to obtain a stable and energetically favorable conformation. During this process, various protonation and tautomeric states were generated at physiological pH 7.4 using Epik to reflect biologically relevant forms of the ligand. Additionally, a thorough stereochemical analysis was performed to identify and select the lowest energy conformer, ensuring accurate representation for subsequent molecular docking studies.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section3\"\u003e \u003ch2\u003e3.2.2. Molecular Docking Active Site Analysis\u003c/h2\u003e \u003cp\u003eTo evaluate the interaction between the synthetic stilbene derivative and the fungal enzyme, molecular docking studies were conducted using the Glide module within the Schr\u0026ouml;dinger suite. The receptor grid was generated via the Receptor Grid Generation tool, with the grid centred on the heme-binding pocket where the native ligand (VT1) is known to bind. Key active site residues such as Tyr118, Gly112, and Arg382 were identified and included in the grid generation due to their crucial role in ligand binding. Docking was performed using a two-tier approach, initially applying the Standard Precision (SP) protocol for broad screening of ligand poses, followed by the Extra Precision (XP) protocol to refine and improve docking accuracy. Ligand binding affinities were ranked based on the GlideScore (GScore), which integrates various energetic and interaction terms. The highest-ranking docking poses were further analysed to identify specific interactions, including hydrogen bonds, π-π stacking, hydrophobic contacts, and any potential metal coordination, providing detailed insight into the binding mode and potential efficacy of the stilbene derivative.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section3\"\u003e \u003ch2\u003e3.2.3. Binding Interaction Analysis\u003c/h2\u003e \u003cp\u003eThe ligand-protein interactions were visualized and analysed using Maestro\u0026rsquo;s Ligand Interaction Diagram tool. This facilitated a detailed examination of the binding mode by clearly illustrating key contacts such as hydrogen bonds, π-π stacking, hydrophobic interactions, and any metal coordination between the synthetic stilbene derivative and the fungal enzyme.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section3\"\u003e \u003ch2\u003e3.2.4. Molecular Docking Results\u003c/h2\u003e \u003cp\u003eThe structure of the selected protein (PDB ID: 4HOE) was validated using a Ramachandran plot generated by PROCHECK RAMPAGE, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e10\u003c/span\u003e [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. The results, summarized in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, indicate that the majority of the amino acid residues are located within the favorable and allowed regions of the plot. This distribution confirms the stereochemical quality and reliability of the protein structure, deeming it suitable for subsequent molecular docking studies.\u003c/p\u003e\u003cp\u003e\u003cimg src=\"data:image/png;base64,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\"\u003e\u003c/p\u003e\n\u003cdiv\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\"\u003e\n \u003ch2\u003e3.2.5. Protein-Ligand Interaction\u003c/h2\u003e\n \u003cp\u003eMolecular docking studies were conducted to evaluate the antifungal potential of hydroxy stilbene against fungal proteins, specifically targeting Candida albicans (PDB ID: 4HOE). The docking results revealed an optimal binding affinity score of -8.0 kcal/mol, indicating a favorable and strong interaction between the hydroxy stilbene derivative and the fungal protein. These findings, illustrated in Fig.\u0026nbsp;11, support the potential efficacy of the compound as an antifungal agent by demonstrating its ability to bind effectively to key fungal targets.\u003c/p\u003e\n \u003cp\u003eThe induced fit docking of the fungal protein 4HOE with 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene produced multiple conformations capable of fitting into the active site pocket, with the lowest energy pose subjected to detailed analysis. Key residues such as Glu32, Phe36, Tyr118, and Ile112 were identified as interacting directly with the Hexahydroxy stilbene, similar to interactions observed with the reference compound UCP111E, as shown on the left. Both hydrogen bonding and π-π interactions contributed to the stabilization of the protein-ligand complex, demonstrating the strong binding capability of 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene within the protein’s active site. The docking scores were calculated as -9.262 kcal/mol for the reference compound and − 11.659 kcal/mol for the Hexahydroxy stilbene, indicating a notably more potent binding affinity of the synthesized compound compared to the reference. Overall, these molecular docking results suggest that hydroxy stilbene possesses significant potential as an effective antifungal agent against Candida albicans. The favorable binding affinities observed align well with previous reports highlighting the antifungal efficacy of stilbene derivatives.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\"\u003e\n \u003ch2\u003e3.3. Density Functional Theory\u003c/h2\u003e\n \u003cp\u003eDensity Functional Theory (DFT) calculations were performed using the JAGUAR program to explore the electronic and structural properties of the ligand. Geometry optimization was conducted employing the B3LYP functional combined with the MIDIX basis set for all atoms, ensuring accurate modeling of molecular orbitals and geometry. To simulate the solvent environment, the polarizable continuum model (PCM) was applied using parameters suitable for DMSO, reflecting realistic experimental conditions. From the optimized molecular structures, key electronic properties such as the energies of the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) were extracted, providing insight into the ligand’s reactivity and stability.\u003c/p\u003e\n \u003cdiv id=\"Sec18\"\u003e\n \u003ch2\u003e\u003cstrong\u003e3.3.1. Density Functional Theory Results\u003c/strong\u003e\u003c/h2\u003e\n \u003cp\u003eThe energy gap (ΔE), defined as the difference between the Highest Occupied Molecular Orbital (HOMO) and the Lowest Unoccupied Molecular Orbital (LUMO), is a vital parameter for assessing the stability and reactivity of a molecule. In this study, DFT calculations were carried out to analyze the electronic properties of the synthesized stilbene derivative, revealing a HOMO-LUMO energy gap of 3.948 eV, as illustrated in Fig.\u0026nbsp;12 and summarized in \u003cstrong\u003eTable\u0026nbsp;2\u003c/strong\u003e. A large energy gap typically indicates high molecular stability, reflecting the significant energy required for electronic excitation. This corresponds to low chemical reactivity, suggesting that the compound is less prone to nucleophilic or electrophilic attack. Such stability is often correlated with enhanced biological activity, as stable molecules tend to form stronger and more specific binding interactions in biological environments. The HOMO is predominantly localized over specific regions of the molecule, highlighting potential sites susceptible to oxidation, whereas the LUMO exhibits delocalization that points to possible electron-accepting sites. This electronic configuration may play a crucial role in the compound’s antifungal activity by influencing its interactions with fungal proteins.\u003c/p\u003e\n \u003cdiv\u003e\n \u003cdiv align=\"left\"\u003e\u003cstrong\u003eTable 2:\u0026nbsp;\u003c/strong\u003eHOMO-LUMO Surface Structures energy gap (ΔE = 3.948 eV) of the stilbene derivative, highlighting its electronic stability and potential reactivity.\u003c/div\u003e\n \u003ctable id=\"Tabc\" border=\"1\"\u003e\n \u003ccolgroup cols=\"4\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCompound Ids\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eE\u003csub\u003eLUMO\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003e(eV)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eE\u003csub\u003eHOMO\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003e(eV)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eΔE\u003csub\u003e(gap)\u003c/sub\u003e = E\u003csub\u003eLUMO\u003c/sub\u003e – E\u003csub\u003eHOMO\u003c/sub\u003e (eV)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-0.997\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-4.945\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.948\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\u003eOverall, the DFT results confirm that the stilbene derivative exhibits high electronic stability, which may enhance its effectiveness as a lead molecule for antifungal drug development.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec19\"\u003e\n \u003ch2\u003e3.4. Biological Evaluation Of 2, 2', 4, 4', 5, 5'-Hexahydroxy Stilbene\u003c/h2\u003e\n \u003cp\u003eScreening for the antifungal target of the selected ligand involved obtaining the three-dimensional (3D) crystal structure of \u003cem\u003eCandida albicans\u003c/em\u003e protein (PDB ID: 4HOE) from the Protein Data Bank. The structure was prepared using the Protein Preparation Wizard in the Schrödinger Suite (Maestro v.2023). During preparation, bond orders were assigned, the hydrogen-bonding network was optimized, and missing hydrogen atoms were added to the structure. Water molecules located more than 5 Å away from the binding pocket were removed to avoid interference, while metal ions critical to maintaining the enzyme’s structural integrity were retained. The protein structure was then energy-minimized using the OPLS4 force field, ensuring proper geometry and stabilization of the enzyme. The active site for molecular docking was defined around the heme-binding region, preserving the native ligand and key catalytic residues to facilitate accurate ligand-receptor interaction studies.\u003c/p\u003e\n \u003cdiv id=\"Sec20\"\u003e\n \u003ch2\u003e3.4.1. Antifungal Activity\u003c/h2\u003e\n \u003cp\u003eThe antifungal potential of the synthesized compounds was evaluated against \u003cem\u003eCandida albicans\u003c/em\u003e, a well-known human pathogenic fungus. The fungal strains were sourced from a certified Microbial Type Culture Collection and Gene Bank (MTCC). Antifungal activity was assessed using the agar dilution method, with compound concentrations ranging from 50 to 200 µg/mL. The minimal inhibitory concentration (MIC) of each test compound was determined following established protocols [10]. Itraconazole, at a dose of 5 grams, was used as a standard reference antifungal agent for comparison.\u003c/p\u003e\n \u003cp\u003eMIC values were specifically determined for yeast isolates that demonstrated sensitivity in the initial disc diffusion assay. The inoculum was prepared from 12-hour-old broth cultures of \u003cem\u003eCandida albicans\u003c/em\u003e. Test compounds were initially dissolved in ethanol and diluted to a maximum concentration of 200 µg/mL. Serial two-fold dilutions were then prepared to achieve concentrations of 50, 100, 150, and 200 µg/mL. The MIC determination was carried out by applying these dilutions onto Potato Dextrose Agar (PDA) plates, designated as P1, followed by incubation and evaluation of fungal growth inhibition.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec21\"\u003e\n \u003ch2\u003e3.4.2. Antifungal Activity by Disc Diffusion Method\u003c/h2\u003e\n \u003cp\u003eThe disc diffusion assay of 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene against \u003cem\u003eCandida albicans\u003c/em\u003e demonstrated a concentration-dependent antifungal effect, with zones of inhibition ranging from 0.8 mm at 200 µg/mL to 18.2 mm at 1000 µg/mL. For comparison, Itraconazole (10 µg/mL) was used as a standard antifungal agent, exhibiting a zone of inhibition of 16.1 mm. These results, illustrated in Fig.\u0026nbsp;13 and detailed in Table\u0026nbsp;3, highlight the promising antifungal activity of the synthesized compound, surpassing the efficacy of the standard drug at higher concentrations.\u003c/p\u003e\n \u003cdiv\u003e\n \u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 3\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003eMinimum inhibition concentration values\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"3\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCompound\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eConcentration\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eCandida albicans\u003c/em\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003e2, 2', 4, 4', 5, 5'-Hexahydroxystilbene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e200 µg/ml\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8 mm\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e400 µg/ml\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.4 mm\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e800 µg/ml\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.1mm\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1000 µg/ml\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18.2 mm\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eItraconazole\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10 µg/ml\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.1mm\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\u003eA comparison study [22] on benzo[4, 5]imidazo[1,2-d][1, 2, 4]triazine derivatives demonstrated significant antifungal activity, primarily attributed to their inhibition of ergosterol biosynthesis, which disrupts fungal cell membrane integrity. In contrast, 2, 2', 4, 4', 5, 5'-Hexahydroxy stilbene exhibits strong antifungal properties through a distinct mechanism. The multiple hydroxyl groups present in Hexahydroxy stilbene play a crucial role in generating reactive oxygen species (ROS), which induce oxidative stress and trigger apoptosis in fungal cells, thereby contributing to the compound’s antifungal efficacy [23]. Besides ROS generation, these hydroxyl groups also interact with the fungal membrane, causing destabilization of the lipid bilayers. This interaction leads to ion leakage and membrane disruption, key factors enhancing its antifungal activity [24]. The dual-action mechanism, involving both oxidative stress and membrane destabilization, is central to the compound’s effectiveness against fungal pathogens. Furthermore, the structure of 2,2',4,4',5,5'-Hexahydroxy stilbene shares similarities with other antifungal agents such as benzo[4, 5]imidazo[1,2-d][1, 2, 4]triazine derivatives [25]. Both compound classes disrupt fungal membranes and inhibit critical biosynthetic pathways. However, the hexahydroxylation of stilbene enhances its hydrophilicity, which improves diffusion into fungal cells, increasing solubility, bioavailability, and ultimately, its antifungal potency.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"4. Conclusion","content":"\u003cp\u003eThis study highlights the strong antifungal potential of the synthetic compound 2,2\u0026prime;,4,4\u0026prime;,5,5\u0026prime;-Hexahydroxy stilbene, highlighting its promise as a novel therapeutic agent. Using an integrated approach\u0026mdash;combining chemical synthesis, spectral characterization, molecular modeling, and biological evaluation\u0026mdash;the research confirms the significance of polyphenolic structures in antifungal activity. Analytical techniques (FTIR, NMR, XRD, and mass spectrometry) validated the compound\u0026rsquo;s structure and purity. Computational docking showed strong binding to fungal protein targets, outperforming standard ligands, while DFT calculations indicated favorable electronic stability. In vitro assays demonstrated potent antifungal activity, comparable or superior to Itraconazole. Its dual mechanism\u0026mdash;ROS generation and membrane disruption\u0026mdash;offers a distinct advantage over conventional drugs. These findings support further investigation of hydroxy stilbene derivatives as promising antifungal candidates amid rising drug resistance.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe take opportunity to thank the University of Mysore and Maulana Azad National Urdu University for giving an opportunity to execute the research.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analyzed during this study are included in this published article [and its Supplementary Information file].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCredit Authorship Contribution Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGopal Krishna Murthy\u003c/strong\u003e: Writing \u0026ndash; review \u0026amp; editing, Writing \u0026ndash; original draft, Methodology, Investigation, Formal analysis.\u0026nbsp;\u003cstrong\u003eRevanasidappa\u003c/strong\u003e: Writing \u0026ndash; review \u0026amp; editing, Writing \u0026ndash; original draft, Visualization, Validation, Methodology, Investigation, Formal analysis.\u0026nbsp;\u003cstrong\u003eShivaprakash.S\u003c/strong\u003e: Writing \u0026ndash; review \u0026amp; editing, Writing \u0026ndash; original draft, Methodology, Investigation.\u0026nbsp;\u003cstrong\u003eVennila Kailasam Natesan\u003c/strong\u003e: Writing \u0026ndash; review \u0026amp; editing, Writing \u0026ndash; original draft, Formal analysis, Data curation.\u0026nbsp;\u003cstrong\u003e\u0026nbsp;Mohammed Saleh Al Ansari\u003c/strong\u003e: Writing \u0026ndash; review \u0026amp; editing.\u0026nbsp;\u003cstrong\u003eSuhail Ahmad\u003c/strong\u003e: Writing \u0026ndash; review \u0026amp; editing, Writing \u0026ndash; original draft, Methodology, Investigation, Formal analysis.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisclosure Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that there are no conflicts of interest regarding the publication of this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo funding was received for the preparation of this article.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eS. Sepehri, M. Khedmati, F. Yousef-Nejad, M. Mahdavi, Medicinal chemistry perspective on the structure\u0026ndash;activity relationship of stilbene derivatives, RSC Adv. D4RA02867H (2024). https://doi.org/10.1039/D4RA02867H\u003c/li\u003e\n\u003cli\u003eD. \u0026Scaron;unjka, \u0026Scaron;. Mechora, An alternative source of biopesticides and improvement in their formulation\u0026mdash;recent advances, Plants (Basel) 11 (22) (2022) 3172. https://doi.org/10.3390/plants11223172\u003c/li\u003e\n\u003cli\u003eE.W.C. Chan, C.W. Wong, Y.H. Tan, J.P.Y. Foo, S.K. Wong, H.T. Chan, Resveratrol and pterostilbene: A comparative overview of their chemistry, biosynthesis, plant sources, and pharmacological properties, J. Appl. Pharm. Sci. 9 (7) (2019) 17\u0026ndash;23. https://doi.org/10.7324/JAPS.2019.90717\u003c/li\u003e\n\u003cli\u003eF. Caruso, L. Mendoza, P. Castro, M. Cotoras, M. Aguirre, B. Matsuhiro, M. Isaacs, M. Rossi, A. Viglianti, Antifungal activity of resveratrol against Botrytis cinerea is improved using 2-furyl derivatives, PLoS One 6 (10) (2011) e25421. https://doi.org/10.1371/journal.pone.0025421\u003c/li\u003e\n\u003cli\u003eL. Mattio, G. Catinella, M. Iriti, L. Vallone, Inhibitory activity of stilbenes against filamentous fungi, Plants (Basel) 10 (4) (2021) 773. https://doi.org/10.3390/plants10040773\u003c/li\u003e\n\u003cli\u003eA. Hammerbacher, S.G. Ralph, J. Bohlmann, T.M. Fenning, J. Gershenzon, A. Schmidt, Biosynthesis of the major tetrahydroxystilbenes in spruce, astringin and isorhapontin, proceeds via resveratrol and is enhanced by fungal infection, Plant Physiol. 157 (3) (2011) 876\u0026ndash;890. https://doi.org/10.1104/pp.111.179325\u003c/li\u003e\n\u003cli\u003eN. Patel, V. Karkhanis, P. Patel, Synthesis and biological evaluation of some piperazine derivatives as anti-inflammatory agents, J. Drug Deliv. Ther. 9 (4-s) (2019) 353\u0026ndash;358. https://doi.org/10.22270/jddt.v9i4-s.3327\u003c/li\u003e\n\u003cli\u003eC.F.G.C. Geraldes, Introduction to infrared and Raman-based biomedical molecular imaging and comparison with other modalities, Molecules 25 (23) (2020) 5547. https://doi.org/10.3390/molecules25235547\u003c/li\u003e\n\u003cli\u003eV. Andrei, A.G.N. Wetie, I. Mihai, C.C. Andrei, Detection of biomedically relevant stilbenes from wines by mass spectrometry, in: J. Bousquet, M. Fountoulakis (Eds.), Advancements of Mass Spectrometry in Biomedical Research, Springer, Cham, 2014, pp. 361\u0026ndash;382. https://doi.org/10.1007/978-3-319-06068-2_17\u003c/li\u003e\n\u003cli\u003eR. Gunawan, A. Nandiyanto, I. Ijost, How to read and interpret \u0026sup1;H-NMR and \u0026sup1;\u0026sup3;C-NMR spectrums, Indones. J. Sci. Technol. 6 (2) (2021) 267\u0026ndash;298. https://doi.org/10.17509/ijost.v6i2.34189\u003c/li\u003e\n\u003cli\u003eJ. O, J. Young, True melting point determination, J. Chem. Educ. 18 (2013) 203\u0026ndash;208. https://doi.org/10.1333/s00897132500a\u003c/li\u003e\n\u003cli\u003eS. Shivaprakash, G.C. Reddy, J.P. Jasinski, S.P. Millikan, C.E. Duff, C. Glidewell, A sterically congested cis-stilbene and its phosphonium salt precursor, Acta Crystallogr. C Struct. Chem. 71 (6) (2015) 479\u0026ndash;484. https://doi.org/10.1107/S2053229615009110\u003c/li\u003e\n\u003cli\u003eS.K. Burley, C. Bhikadiya, C. Bi, et al., RCSB Protein Data Bank (RCSB.org): delivery of experimentally-determined PDB structures alongside one million computed structure models of proteins from artificial intelligence/machine learning, Nucleic Acids Res. 51 (D1) (2023) D488\u0026ndash;D508. https://doi.org/10.1093/nar/gkac1077\u003c/li\u003e\n\u003cli\u003eT. Egawa, S.-R. Yeh, D. Rousseau, Redox-controlled proton gating in bovine cytochrome c oxidase, PLoS One 8 (2013) e63669. https://doi.org/10.1371/journal.pone.0063669\u003c/li\u003e\n\u003cli\u003eW. Tian, C. Chen, X. Lei, J. Zhao, Y. Li, Q. Wang, CASTp 3.0: Computed atlas of surface topography of proteins, Nucleic Acids Res. 46 (W1) (2018) W363\u0026ndash;W367. https://doi.org/10.1093/nar/gky473\u003c/li\u003e\n\u003cli\u003eC. Colovos, T.O. Yeates, Verification of protein structures: patterns of nonbonded atomic interactions, Protein Sci. 2 (9) (1993) 1511\u0026ndash;1519. https://doi.org/10.1002/pro.5560020916\u003c/li\u003e\n\u003cli\u003eO. Trott, A.J. Olson, AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading, J. Comput. Chem. 31 (2) (2010) 455\u0026ndash;461. https://doi.org/10.1002/jcc.21334\u003c/li\u003e\n\u003cli\u003eA.C. Wallace, R.A. Laskowski, J.M. Thornton, LIGPLOT: A program to generate schematic diagrams of protein-ligand interactions, Protein Eng. 8 (2) (1995) 127\u0026ndash;134. https://doi.org/10.1093/protein/8.2.127\u003c/li\u003e\n\u003cli\u003eS. Forli, R. Huey, M.E. Pique, M.F. Sanner, D.S. Goodsell, A.J. Olson, Computational protein\u0026ndash;ligand docking and virtual drug screening with the AutoDock suite, Nat. Protoc. 11 (5) (2016) 905\u0026ndash;919. https://doi.org/10.1038/nprot.2016.051\u003c/li\u003e\n\u003cli\u003eG. Li, X. Ma, L. Deng, X. Zhao, Y. Wei, Z. Gao, J. Jia, J. Xu, C. Sun, Fresh garlic extract enhances the antimicrobial activities of antibiotics on resistant strains in vitro, J. Infect. Microbiol. 10 (2025) 14814. https://doi.org/10.5812/jjm.14814\u003c/li\u003e\n\u003cli\u003eA.V. Snezhkina, A.V. Kudryavtseva, O.L. Kardymon, M.V. Savvateeva, N.V. Melnikova, G.S. Krasnov, A.A. Dmitriev, ROS generation and antioxidant defense systems in normal and malignant cells, Oxid. Med. Cell. Longev. 2019 (2019) 6175804. https://doi.org/10.1155/2019/6175804\u003c/li\u003e\n\u003cli\u003eP. Nakhaei, R. Margiana, D.O. Bokov, W.K. Abdelbasset, M.A. Jadidi Kouhbanani, R.S. Varma, F. Marofi, M. Jarahian, N. Beheshtkhoo, Liposomes: structure, biomedical applications, and stability parameters with emphasis on cholesterol, Front. Bioeng. Biotechnol. 9 (2021) 705886. https://doi.org/10.3389/fbioe.2021.705886\u003c/li\u003e\n\u003cli\u003eT. Kon, N. Nemoto, T. Oshima, A. Yamagishi, Effects of a squalene epoxidase inhibitor, terbinafine, on ether lipid biosyntheses in a thermoacidophilic archaeon, Thermoplasma acidophilum, J. Bacteriol. 184 (5) (2002) 1395\u0026ndash;1401. https://doi.org/10.1128/JB.184.5.1395-1401.2002\u003c/li\u003e\n\u003cli\u003eM. Krysa, M. Szymańska-Chargot, A. Zdunek, FT-IR and FT-Raman fingerprints of flavonoids \u0026ndash; a review, Food Chem. 393 (2022) 133430. https://doi.org/10.1016/j.foodchem.2022.133430\u003c/li\u003e\n\u003cli\u003eK. Jomova, R. Raptova, S.Y. Alomar, S.H. Alwasel, E. Nepovimova, K. Kuca, M. Valko, Reactive oxygen species, toxicity, oxidative stress, and antioxidants: chronic diseases and aging, Arch. Toxicol. 97 (10) (2023) 2499\u0026ndash;2574. https://doi.org/10.1007/s00204-023-03562-9\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"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":"Hydroxy stilbene, Antifungal, Molecular docking, DFT","lastPublishedDoi":"10.21203/rs.3.rs-6755724/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6755724/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eStilbenes are a large class of plant-derived secondary metabolites belonging to the polyphenol family and are widely distributed in foods such as grapes, peanuts, and other dietary sources. They exist in two isomeric forms—\u003cem\u003etrans\u003c/em\u003e-stilbene (E-form) and \u003cem\u003ecis\u003c/em\u003e-stilbene (Z-form)—with several natural derivatives, including resveratrol, pterostilbene, and combretastatin A-4 (a polymethoxylated \u003cem\u003ecis\u003c/em\u003e-stilbene), exhibiting a broad spectrum of biological activities such as antimicrobial, anticancer, anti-inflammatory, cardioprotective, neuroprotective, and antidiabetic effects. The \u003cem\u003ecis\u003c/em\u003e-stilbene configuration, in particular, offers distinct structural and electronic characteristics that make it highly valuable in the fields of photochemistry, materials science, and medicinal chemistry. In the present work, we report the synthesis of \u003cem\u003ecis\u003c/em\u003e-stilbene derivative, (Z)-2,2′,4,4′,5,5′-hexahydroxy stilbene, achieved for the first time in a single-step demethylation reaction from its precursor, (Z)-2,2′,4,4′,5,5′-hexamethoxy stilbene. The structure and purity of the synthesized compound were confirmed using Fourier-transform infrared (FTIR) spectroscopy, mass spectrometry (MS), and nuclear magnetic resonance (NMR) spectroscopy, including both ¹H and ¹³C NMR. The Z-configuration and detailed molecular structure were further validated through single-crystal X-ray diffraction analysis. In addition to structural characterization, density functional theory (DFT) studies were conducted to understand the molecule’s electronic properties, which complemented the experimental data.\u003c/p\u003e","manuscriptTitle":"Design, Synthesis, Structural, and Computational Studies of (Z)-2,2′,4,4′,5,5′-Hexahydroxy Stilbene as a Potential Antifungal Agent","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-17 15:59:53","doi":"10.21203/rs.3.rs-6755724/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":"b0654c1b-a2e7-4825-8643-778960075ba9","owner":[],"postedDate":"June 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":50136005,"name":"Biological sciences/Biochemistry"},{"id":50136006,"name":"Physical sciences/Chemistry"}],"tags":[],"updatedAt":"2025-06-26T17:38:27+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-17 15:59:53","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6755724","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6755724","identity":"rs-6755724","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}