Bioactive triterpenoids isolated and characterized from Ficus sur (Forrsk) stem bark extract demonstrate inhibitory activity against benign prostatic hyperplasia protein targets using in silico study

Bioactive triterpenoids isolated and characterized from Ficus sur (Forrsk) stem bark extract demonstrate inhibitory activity against benign prostatic hyperplasia protein targets using in silico study | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Bioactive triterpenoids isolated and characterized from Ficus sur (Forrsk) stem bark extract demonstrate inhibitory activity against benign prostatic hyperplasia protein targets using in silico study Uchenna Benjamin Okeke, Patrick Igbinaduwa, Ibrahim Oluwatobi Kehinde, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8786768/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 17 You are reading this latest preprint version Abstract Plants possess an abundant reservoir of compounds that can be harnessed and repurposed as therapeutic agents for the management of existing and emerging disease conditions, such as benign prostatic hyperplasia (BPH). This study aimed to isolate and characterize bioactive phytochemicals from the ethyl acetate fraction of the Ficus sur hydroethanol stem bark extract (FSSE) and to carry out in silico evaluation of their activity against key protein targets implicated in BPH pathogenesis. Ficus sur stem bark was collected from its natural habitat, authenticated, extracted, and partitioned into different solvent fractions (dichloromethane, ethyl acetate, and n- butanol). Sequential open column chromatography and preparatory thin layer chromatography of the ethyl acetate fraction led to the isolation and purification of compounds, which were characterized by spectroscopic analysis (UV, FTIR, 1 H and 13 C-NMR, and DEPTQ experiments). An In-silico molecular docking study was conducted using Autodock Vina, while ADMETlab 3.0 was employed to predict the physicochemical properties and drug-likeness of the isolated compounds. Two triterpenoids, α-amyrin acetate (110 mg) and β-sitosterol (340 mg), from the eight isolated compounds were characterized. In silico analyses indicated that both compounds exhibited binding affinities and physicochemical profiles comparable to those of the co-crystallized ligand finasteride across the four molecular targets implicated in benign prostatic hyperplasia. This gave credence to the medicinal richness of this plant and its use in folklore for urinary-related disorders such as BPH. It has been established in this study that F. sur stem bark extract is rich in bioactive compounds that can be explored for the discovery of leads for developing new drug candidates, especially for BPH management. Ficus sur amyrin acetate sitosterol prostatic hyperplasia triterpenoid 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 Figure 14 Figure 15 1 Introduction The search for chemical entities from natural products for the management of emerging and complex disease conditions has continued to experience a quantum leap, creating a basis for more extensive scientific exploration of diverse lead structures that serve as templates for developing new pharmaceuticals. The growing interest in herbal medicines is largely driven by the perception that they are safer and more affordable than synthetic drugs, which are often expensive and associated with adverse side effects. Studies have revealed that various phytochemicals, such as alkaloids, flavonoids, phenolics, terpenoids, and saponins, found in medicinal plants can trigger a range of biological activities in living organisms, including antioxidant, anti-inflammatory, antibacterial, antiviral, and antitumor effects (Zhang et al., 2024 ). In vitro and in vivo studies validated these hypotheses that plant-derived compounds act through multiple pathways (Latif and Nawaz, 2025 ), including inhibition of 5α-reductase, a key enzyme in benign prostatic hyperplasia and prostate cancer pathogenesis, due to their structural similarity to endogenous receptor ligands (Nguyen et al., 2024 ). Benign prostatic hyperplasia (BPH), a nonmalignant enlargement of the prostate causing lower urinary tract symptoms (LUTS), has become a significant health issue among aging men worldwide (Speakman et al., 2015 ). Due to the bothersome side effects linked to conventional medications, the demand for herbal treatments for BPH management has greatly increased. Herbal remedies, some available in formulated dosage forms, have demonstrated effectiveness in treating BPH, cancer chemotherapy, and chemoprophylaxis. For example, herbal preparations containing either single or combined extracts of Saw palmetto, Serenoa repens, Pygeum africanum, Urtica dioica, Cucurbita pepo , and β-sitosterol from Hypoxis rooper (African star grass) help manage BPH (Csikós et al., 2021 ). Biological activity-guided isolation techniques have produced important therapeutic agents for diseases like cancer, Alzheimer’s disease, infections, malaria, and diabetes (Atanasov et al., 2021 ). Advances in computer-aided drug design have further sped up the discovery and development of plant-derived drugs, making them increasingly promising. To achieve significant improvements and increase cost-efficiency in drug development, employing computational methods has become essential. Typically, after identifying and selecting disease-related targets, various computational tools perform key drug discovery steps, including target identification (such as an enzyme or protein receptor), modifying drug-like compounds, and optimizing these compounds to develop effective drug candidates (Duffy et al., 2012 ). Ficus sur , also known as broom cluster fig or cape fig, belongs to the genus Moraceae and is widely distributed in several tropical African countries. The plant, due to its high medicinal richness, has been used in ethnomedicine to treat various disease conditions such as infections, tumors, pains, and urinary disorders (Sieniawska et al., 2022 ). Several phytochemicals, such as triterpenoids, polyphenols, and fatty acids, have been isolated from different parts of F. sur in preceding studies (Ogunlaja, 2022; Sieniawska et al., 2022 ). Several phytosterol (β-sitosterol, stigmasterol), oleanane-type (12-oleanen-3-yl acetate, oleanen-3-yl acetate), and ursine-type (urs-12-en-24-oic acid, α-amyrin, 3-acetoxy-α-amyrin) triterpenoids have been isolated from the latex, leaf, and stem bark of F. sur (Nwanisobi et al., 2021 ; Ogunlaja et al., 2022 ; Sieniawska et al., 2022 ; Mouelle et al., 2025 ). Okeke et al ( 2026 ) reported the presence of gamma-sitosterol from the F. sur hydroethanolic stem bark extract. However, there is a paucity of scientific validation of the efficacy of these phytochemicals on benign prostatic hyperplasia. This work reports herein the isolation and characterization of two triterpenoids, α-amyrin acetate and β-sitosterol, from the ethyl acetate fraction of the hydroethanolic stem bark of F. sur and in silico evaluation of their ameliorating activity against protein targets implicated in BPH pathogenesis. 2 Materials and Methods 2.1 Collection, extraction, and fractionation of plant material F. sur stem bark was identified and collected from its natural habitat in Ibadan, Oyo State, Nigeria (Fig. 1 ), by Dr. Akinniyi Samuel Odewo, a Taxonomist at the Forest Research Institute of Nigeria (FRIN), Ibadan, with compliance to WHO guidelines on good agricultural and collection practices for medicinal plants (World Health Organization, 2003). A herbarium specimen of the plant material (FHI113940) was deposited in the facility. The powdered stem bark (5 kg) was extracted by maceration with 80% ethanol at room temperature for 72 hours. The solution was filtered using Whatman No. 4 filter paper, and the filtrate was concentrated to dryness under vacuum at 50°C using a rotary evaporator to afford F. sur hydroethanol stem bark extract (FSSE) (yield: 286 g, 5.72% w / w ). Employing the modified Kupchan method of solvent partitioning as described by Van-Wagenen et al. ( 1993 ), 250 g of the extract initially suspended in 200 mL of distilled water was transferred completely into a 2000 mL separating funnel and successively partitioned in the order: n -hexane (HEX), dichloromethane (DCM), ethyl acetate (EtOAc), and n -butanol (BuOH). The various fractions collected were concentrated under vacuum in a rotary evaporator to obtain the concentrates, which were then air-dried and weighed. 2.2 Isolation and purification of compounds Firstly, the ethyl acetate fraction (40.00 g) of FSSE was adsorbed onto silica gel (20.00 g, 70–230 mesh size) in a mortar to form a slurry, which was then air-dried and loaded into a 50 x 10 cm glass column packed with 500 g of silica gel (70–230 mesh). Dichloromethane (100%) was used to pack the column before loading the sample. The column was eluted with gradient solvent systems of increasing polarity in the order: n -hexane, dichloromethane, and ethyl acetate. A total of 32 fractions were collected (250 mL each) until the compounds were completely eluted from the column. The fractions were monitored by thin-layer chromatography (TLC), and those with similar TLC profiles were pooled and reduced in vacuo at 45°C to obtain 7 fractions (FSE-1 to FSE-7). Fractions FSE-1 (1–5), FSE-2 (6–9), and FSE-4 (13–14) were subjected to a series of open column chromatography (CC) and preparative thin-layer chromatography (PTLC) using gradient solvent systems. The various sub-fractions obtained were monitored with TLC to confirm the presence of a single spot. The isolated compounds (C1 – C8) (Fig. 2 ) were identified by TLC and melting point determination. The structures of the compounds were elucidated through spectral analysis, including ultraviolet (UV), Fourier Transform Infrared (FTIR), ¹H and ¹³C NMR, and DEPTQ spectroscopic experiments, and by comparing the spectral data with literature values. 2.3 Molecular docking of isolated compounds The chemical structures of the isolated and characterized compounds were drawn using ChemDraw and converted to 3D structures. The compounds were optimized by correcting stereochemistry, removing duplicates, and adjusting protonation states to physiological pH (Omotuyi et al., 2021 ). Molecular docking was performed using AutoDock Vina (version 1.2.3) to predict the binding affinities and interaction modes of the compounds with the BPH-related protein targets: 5α-reductase 2 (5αR2; PDB ID: 7BW1), α 1A -adrenoceptor (ADRA1A; PDB ID: 7YMJ), phosphodiesterase-5 (PDE5; PDB ID: 2H42), and muscarinic M3 receptor (PDB ID: 5ZHP) (Fig. 3 ) (Eberhardt et al., 2021 ; Trott and Olson, 2010 ). Before docking, the prepared protein and ligand structures were converted into the PDBQT format using AutoDock Tools (ADT, version 1.5.7). Docking calculations were carried out for each protein target using the prepared ligands within the defined grid box. For each ligand, AutoDock Vina generated multiple binding poses ranked by predicted binding affinity (kcal/mol). Docking results were visualized and analyzed for protein–ligand interactions using UCSF Chimera and BIOVIA Discovery Studio Visualizer (Pettersen et al. , 2004). 2.4 Drug-likeness and in-silico ADMET prediction analysis of isolated compounds ADMETlab 3.0 Web tool ( https://admetlab3.scbdd.com/ ) was used to predict the physicochemical properties, toxicity profile, and drug-likeness nature of the isolated compounds. The drug-likeness was evaluated based on the Lipinski rule, which is related to the compounds' predicted absorption, distribution, metabolism, and excretion (ADME) properties. Compounds with multiple violations are generally less likely to demonstrate favourable in vivo activity. Topological polar surface area (TPSA), another critical pharmacokinetic parameter, was also examined, as it predicts membrane permeability of polar atoms; optimal passive absorption typically falls within the range of 20–140 Ų (Fu et al., 2024 ). 3 Results and Discussion Open column chromatography (CC) and PTLC separation methods on Silica gel afforded the isolation of majorly semi-polar constituents (C1 to C8) from the ethyl acetate fraction of FSSE. The structure of the isolated compounds was confirmed using various spectral analysis techniques like ¹H NMR, ¹³C NMR, DEPTQ, and comparison of the spectra data with those in the literature. 3.1 Physical characteristics and identification of isolated compounds Compound C5 α-Amyrin acetate. Isolated and purified as a 240 mg white solid, soluble in 99% chloroform, with a melting point of 225–227°C and a positive Liebermann-Buchard test for steroids. Spectral Analysis UV-vis spectrum (CHCl 3 solvent) recorded λmax at 227 nm in chloroform. The FTIR spectrum showed absorption bands (in cm − 1 ) at 2952.1, 2911.1, 1733.2, 1244.9, and 1457.4, respectively. This reflects the presence of key functional groups in the compound. Pansare et al. ( 2021 ) reported a similar FTIR absorbance pattern for α-amyrin acetate. The 1 H NMR spectrum (CDCl 3 solvent; 600 MHz NMR spectrometer) of C5 shown in Fig. 4 , revealed various proton peaks as follows (δ in ppm): 5.18 (1H, t , H-12), 4.51 (1H, t , H-3), 2.30 (3H, s , H-2'), 2.01 (1H, m , H-20), 1.90 (2H, t , H-11), 1.63 (2H, m , H-2), 1. 61 (1H, t , H-9), 1.53 (2H, t , H-6). Notably, the deshielded vinyl protons H-12 and H-3 peaks appeared downfield as a triplet at 5.15 ppm and a doublet of doublets at 4.50, respectively. A sharp, intense singlet at 2.10 ppm corresponds to the three methyl protons of an acetate group, confirming the presence of an acetate moiety. These spectral features are consistent with an ursane-type pentacyclic triterpenoid, in contrast to the oleanane-type (e.g., β-amyrin), with a H-12 proton resonating near 5.18 ppm. Eight methyl signals are clustered in a crowded upfield region between 0.65 and 1.20. The 13 C NMR and DEPTQ spectra in Figs. 5 and 6 revealed a total of 32 carbon signals, categorized as eight methyl (-CH 3 ), nine methylene (-CH 2 ), seven methine (-CH), and seven quaternary (–C-) carbons, detailed as follows (CDCl 3 solvent; 600 MHz NMR spectrometer): δ ppm 170.95 (C-1'), 139.64 (C-13), 124.34 (C-12), 80.97 (C-3), 59.10 (C-18), 55.30 (C-5), 47.67 (C-9), 42.10 (C-14), 41.74 (C-8), 40.06 (C-22), 39.83 (C-1). The chemical shift values presented in Table 1 align closely with those reported for α-amyrin acetate by Okoye et al. ( 2014 ), indicating concordance between the observed and literature NMR spectra. A characteristic signal at 170.92 ppm indicates a carbonyl carbon (C = O) of an ester. Peaks observed downfield at 124.34 and 139.64 ppm correspond to the olefinic C-12 methine and C-13 quaternary carbons, respectively. The methine carbons C-18 and C-19 appeared downfield at 59.10 and 39.84 ppm. These assignments further support the characterization of the compound as an ursane-type triterpenoid, distinguishing it from the oleanane framework, which typically shows C-12 and C-13 resonance peaks near 121 and 145 ppm, and C-18 and C-19 signals nearly overlapping around 47–47.5 ppm (Okoye et al., 2014 ; Viet et al., 2021 ). 1 H- 1 H COSY of C5 (Fig. 11 ) revealed cross-peaks indicating coupling between neighboring protons, particularly H-2 / H-3 (1.6 and 4.55 ppm), H-11 / H-12 (1.9 and 5.15 ppm), and H-12 / H-18 (5.15 and 1.2 ppm). Table 1 13 C-NMR and 1 H-NMR data of C5 and α-amyrin acetate from literature Position Chemical shift, δ (ppm) value Compound C5 (δ ppm) Chemical shift, δ (ppm) value Compound C5 (δ ppm) α-amyrin acetate (Okoye et al., 2014 ) 1 H 13 C 1 H 13 C 13 C NMR 1 2 1.63 (m) 3 4.55 (dd) 80.95 4.56 (dd) 80.97 81.18 4 5 55.30 55.30 55.46 6 1.53 (t) 7 8 39.67 39.83 9 1.61 (t) 47.68 47.68 47.84 10 11 1.90 (t) 1.98 12 5.15 (t) 124.34 5.18 (t) 124.34 124.5 13 139.64 139.64 139.8 14 42.10 42.10 42.10 15 16 17 18 59.10 59.10 59.26 19 39.84 20 2.01 (m) 21 22 23 24 25 26 27 28 29 30 1' 170.92 170.95 171.53 2' 2.1 Chemical shift values expressed as parts per million (ppm); s- singlet, d- doublet, t- triplet, dd- doublet of a doublet, m- multiplet Compound C8 β-Sitosterol. Isolated and purified as a 341 mg pale yellow crystalline solid, soluble in 99% chloroform, with a melting point of 136–138°C, and a positive Liebermann-Buchard test for steroids. Spectral analysis : UV-vis spectrum (CHCl 3 solvent) recorded λmax at 260 nm, principally attributed to a single isolated double bond and the solvent effect (bathochromic shift) of chloroform. 1 H NMR spectrum (CDCl 3 solvent, 600 MHz NMR) revealed various proton peaks at different chemical shifts (δ in ppm) as follows: 5.3 (1H, s , H-6), 3.48 (1H, m , H-3), 2.2 (2H, m , H-2), 2.2 (2H, m , H-4), 1.9 (2H, t , H-7) (Fig. 7 ). Table 2 reveals chemical shift values that correspond closely with the β-sitosterol NMR spectrum documented by Ododo et al. ( 2016 ) and Javed et al. ( 2021 ), confirming NMR spectral consistency between the observed and literature values. A distinct multiplet peak at 5.36 ppm corresponds to the olefinic proton on carbon C-6. The deshielded hydroxyl group proton on carbon C-3 appears as a doublet of doublets at 3.49 ppm. 13 C NMR and DEPTQ spectra of compound C5 in Figs. 8 and 9 , revealed a total of 29 carbon signals as six methyl (-CH 3 ), eleven methylene (-CH 2 ), nine methine (-CH) and three quaternary (-C-) carbons, detailed as follows (600 MHz, in CDCl 3 solvent) δ ppm: 140.76 (C-5), 129.30 (C-6), 71.81 (C-3), 56.89 (C-17), 56.79 (C-14), 50.1 (C-9), 45.87 (C-24), 42.33 (C-13), 42.3 (C-4), 40.47 (C-12), 37.20 (C-1), 36.51(C-10), 36.15 (C-20). 1 H- 1 H COSY revealed cross-peaks indicating correlations between neighboring protons in compound C8 (Fig. 12 ), particularly H-2 / H-3 (2.3 and 3.49 ppm), H-3 / H-4 (3.39 ppm and 2.29 ppm), and H-6 / H-7 (5.35 and 1.98 ppm). Table 2 13 C-NMR and 1 H-NMR data of C8 and β-sitosterol from literature Position Compound C8 chemical shift, δ (ppm) β-Sitosterol (Javed et al., 2021 ) β-Sitosterol (Ododo et al., 2016 ) β-Sitosterol (Javed et al., 2021 ) β-Sitosterol (Ododo et al., 2016 ) 1 H 13 C 1 H NMR 1 H NMR 13 C NMR 13 C NMR 1 37.27 1.46 (m) 36.8 37.28 2 2.3 (m) 1.56 (m) 31.2 31.69 3 3.49 (m) 71.81 3.45 3.54 (m) 72.2 71.82 4 2.29 (m) 42.23 (t) 2.32 (m) 42.4 42.33 5 - 140.76 - - 141.2 140.77 6 5.35 (t) 121.71 5.41 5.37 (t) 122.1 121.73 7 1.98 (t) 31.88 2.04 (m) 31.8 31.93 8 31.63 1.69 (m) 31.63 31.93 9 50.19 1.55 (m) 51.9 50.16 10 36.15 - 35.8 36.51 11 1.52 (m) 22.8 21.11 12 40.47 1.51 (m) 40.6 39.80 13 39.80 - 41.8 42.34 14 56.88 1.50 (m) 56.8 56.79 15 24.31 1.58 (m) 24.8 24.33 16 1.85 (m) 28.6 28.27 17 56.09 1.45 (m) 56.2 56.08 18 0.68 0.70 (s) 12.6 11.89 19 1.03 1.03 (s) 20.6 19.42 20 33.97 1.60 (m) 34.4 36.17 21 0.92 0.94 (d) 18.9 18.84 22 31.92 0.93 (m) 32.2 33.98 23 26.1 1.15 (m) 25.4 26.11 24 45.87 1.38 (m) 46.4 45.86 25 29.70 1.57 (m) 30.4 29.19 26 0.86 0.84 (d) 21.2 19.84 27 0.82 0.86 (d) 19.81 19.06 28 23.09 1.10 (m) 23.8 23.10 29 0.89 0.82 (t) 12.8 12.01 C-3 OH - 1.98 (s) - - Chemical shift values expressed as parts per million (ppm); s- singlet, d- doublet, t- triplet, m- multiplet 3.2 Molecular docking analysis of isolated bioactive compounds against BPH targets The docking scores, representing the predicted binding free energies (kcal/mol) of α-amyrin acetate and β-sitosterol, isolated from the ethyl acetate fraction of FSSE, against key therapeutic targets implicated in benign prostatic hyperplasia (BPH), are presented in Table 3 . Table 3 Molecular docking scores (kcal/mol) of α-amyrin acetate and β-sitosterol against BPH targets Compounds 5α reductase 2 α 1A - adrenoceptor Phosphodiesterase-5 Muscarinic M3 PDB ID: 7BW1 PDB ID: 7YMJ PDB ID: 2H42 PDB ID: 5ZHP α-Amyrin acetate (C5) -10.2 -10.9 -9.8 -8.0 β-Sitosterol (C8) -10.7 -8.0 -6.8 -7.7 PDB – Protein Data Bank 3.3 Molecular interaction analysis of Ficus sur bioactive compounds with BPH target proteins Figure 13 and 14 depicted the two-dimensional (2D) interaction profiles of the isolated compounds in Fig. 10 , docked against the four major therapeutic targets of BPH. The visualized interactions highlight a combination of Van der Waals forces, π–π stacking, π–alkyl, alkyl, and hydrogen bonding, which collectively contribute to the stability and specificity of the ligand–receptor complexes, corroborating the docking energy results in Table 3 . α-Amyrin acetate complexes displayed dense networks of hydrophobic and aromatic interactions across all receptor pockets, indicating a strong binding affinity and multi-target potential. Within the 5α-reductase 2 complex (7BW1), α-amyrin acetate established extensive π–alkyl and alkyl contacts with residues such as Phe118, Arg114, Leu23, and Val24, while also forming Van der Waals interactions with Ala26 and Gly115, stabilizing the ligand within the hydrophobic catalytic channel of the enzyme. In the α 1A -adrenoceptor complex (7YMJ), the compound interacted prominently with Trp102, Tyr176, and Phe312 through π–π and hydrophobic contacts, suggesting a strong occupation of the receptor’s transmembrane binding site that could contribute to adrenergic antagonism. Similarly, in the phosphodiesterase-5 (2H42) complex, α-amyrin acetate formed stabilizing interactions with Phe820, Met816, and Ala823, complemented by π–alkyl contacts that mimic known inhibitor binding patterns in the cGMP catalytic domain. For the muscarinic M3 receptor (5ZHP), α-amyrin acetate maintained π–π and Van der Waals interactions with residues Phe437, Arg109, and Asn107, suggesting that it may also influence bladder contractility through muscarinic receptor modulation. These interactions explain the consistently high docking energies observed (− 10.9 to − 8.0 kcal/mol) and reinforce the compound’s potential as a multi-target modulator in BPH. β-Sitosterol also exhibited favorable interaction patterns with all four BPH targets. In the 5α-reductase binding pocket, β-sitosterol engaged in π–π stacking and alkyl interactions with key hydrophobic residues, including Phe118, Tyr176, and Leu23, which are crucial for enzyme inhibition and suppression of dihydrotestosterone biosynthesis. The α 1A -adrenoceptor complex revealed π–alkyl interactions with Phe312 and Trp102, along with hydrogen bonding to Tyr196, which collectively suggest strong antagonistic behavior similar to known α1-blockers. Within the phosphodiesterase-5 complex, β-sitosterol interacted mainly with hydrophobic residues Phe820, Met816, and Val826, while the M3 receptor complex exhibited π–π stacking with Phe437 and hydrophobic contacts with Ile424 and Val426. The abundance of hydrophobic and aromatic contacts indicates that β-sitosterol’s lipophilic nature allows it to efficiently stabilize within receptor pockets, consistent with its recognized pharmacological role as a 5α-reductase inhibitor and prostate health modulator. The molecular interaction analysis confirms that the investigated phytoconstituents, α-amyrin acetate and β-sitosterol, exhibit strong and complementary binding profiles across multiple BPH-related targets. Their interactions are dominated by π–π and hydrophobic forces involving catalytically and structurally relevant residues, which not only stabilize the complexes but also suggest a synergistic multi-target therapeutic mechanism. This molecular evidence supports the hypothesis that the bioactive constituents of Ficus sur exert anti-BPH activity through concurrent inhibition of androgenic (5α-reductase), adrenergic (α 1A− adrenoceptor), phosphodiesterase (PDE-5), and muscarinic receptor pathways, offering a mechanistic rationale for the traditional use of Ficus sur in prostate and urinary-related disorders. 3.4 Drug-Likeness and ADMET Properties of Isolated Compounds The drug-likeness and pharmacokinetic profile of α-amyrin acetate (C5) and β-sitosterol (C8) isolated from ethyl acetate fraction of FSSE were evaluated using the ADMETlab 3.0 web interface, with reference to Lipinski’s rule of five (molecular mass (< 500 Da), lipophilicity (Log P ≤ 5), hydrogen bond donors (HBD ≤ 5), and hydrogen bond acceptors (HBA ≤ 10) (Lipinski et al., 2001 ), along with additional pharmacokinetic descriptors. Table 3 summarises the key parameters supporting the suitability of the isolated compounds for biological evaluation, including their predicted physicochemical properties, lipophilicity, water solubility, pharmacokinetics, and overall drug-likeness. α-Amyrin acetate and β-sitosterol demonstrated favourable oral bioavailability, obeying Lipinski’s rule of 5. The ADMETLab output for the two compounds showed a coloured zone that represents the suitable physicochemical space for oral bioavailability, based on key physicochemical descriptors. Table 3 presents the critical parameters supporting Compounds C5 and C8 as candidates for biological evaluation. These features further suggest that both compounds are likely to demonstrate good oral bioavailability, which reinforces the potential of the compounds as therapeutic candidates for the treatment of BPH and other bladder disorders. Table 3 ADMET physicochemical properties of ligands with the best binding affinities Property Meaning Optimal Value α-Amyrin acetate β-Sitosterol MW Molecular weight 150 mg/mol ˂ MW ≤ 500 g/mol 468.4 414.39 LogP Lipophilicity 0–3 log mol/L *5.799 *8.004 LogS Aqueous solubility -4–0.5 log mol/L -6.519 -7.221 LogD Distribution coefficient (pH 7.4) 1–3 log mol/L (pH 7.4) 4.211 5.37 TPSA Topological polar surface area 0 ~ 140A 2 26.3 20.23 nHA Number of hydrogen bond acceptors 0 ~ 12 2.0 1.0 nHD Number of hydrogen bond donors 0 ~ 7 0.0 1.0 nRot Number of rotatable bonds 0 ~ 11 2.0 6.0 nRing Number of rings 0 ~ 6 5.0 4.0 nRig Number of rigid bonds 0 ~ 30 27.0 20.0 MaxRing Number of atoms in the biggest ring 0–18 22.0 17.0 fCHar Formal charge -4 ~ 4 0.0 0.0 nHet Number of heteroatoms 1 ~ 15 2.0 1.0 NLRV Number of Lipinski’s violations 1 1 * - LRV : Lipinski’s rule violation; NLRV : Number of Lipinski’s rule violations 4 Conclusion Open column chromatography, preparative thin-layer chromatography, and instrumental analytical techniques such as UV, FTIR, 1 H and 13 C NMR, resulted in the isolation of an ursane-type pentacyclic triterpenoid, α-Amyrin acetate (C5), and a phytosterol, β-sitosterol (C8). Both compounds (α-amyrin acetate and β-sitosterol) exhibited ameliorative effects against BPH through molecular docking on BPH protein targets, with β-sitosterol displaying binding scores comparable to those of the standard BPH drug, finasteride. This study highlights the diversity of this plant, validation of its traditional use and potential as a source of lead compounds for developing therapeutics to treat BPH and other urinary-related conditions, and contribution to improvement in public health strategies for managing BPH, especially in developing regions where access to standard therapies may be limited. Declarations Acknowledgments The authors acknowledge the contributions of the Faculty of Pharmacy, University of Benin, Nigeria, and Afe Babalola University, Ado-Ekiti, Nigeria, for providing the facilities for conducting this research, and also the School of Chemistry and Physics, University of KwaZulu-Natal, Durban, South Africa, for providing the facilities for the NMR experiments. Author Contributions Uchenna Benjamin Okeke: Research concept development, data collection, analysis, manuscript draft preparation, review, and editing. Patrick Igbinaduwa: Research concept development, data analysis, and critical reading of manuscript. Vuyisa Mzozoyana: NMR analysis and data collection. Ibrahim Oluwatobi Kehinde: Bioinformatics analysis, article draft review, and editing. Funding The authors declare that no funds, grants, or other support were received during the research work and preparation of this manuscript. Data Availability The authors declare that the data supporting the findings of this study are available within the paper. Should any raw data files be needed in another format, the corresponding author, upon reasonable request and with the consent of the other authors of this research, shall make them available. Consent for publication The authors declare no conflict of interest. We confirm the publication of this article. Competing Interests The authors declare that they have no competing interests. Ethics and consent to participate Permissions or licences for plant collection and animal use are not applicable in this study, as the plant is common and non-endangered. Ficus sur herbarium specimen (Voucher number: FHI113940) was deposited at Forest Herbarium Ibadan (FHI), located in the Taxonomy Section of the Department of Forest Conservation and Protection, Forestry Research Institute of Nigeria (FRIN), Ibadan. The plant identifier and collector at FRIN is Dr. Akinniyi Samuel Odewo. 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Phytochemical Insights into Ficus sur Extracts and Their Biological Activity. Molecules 2022;27(6):1863. doi: 10.3390/molecules27061863. Speakman M, Kirby R, Doyle S, Ioannou C. Burden of male lower urinary tract symptoms (LUTS) suggestive of benign prostatic hyperplasia (BPH) - focus on the UK. BJU Int. 2015;115(4):508-19. https://doi.org/10.1111/bju.12745. Trott O, Olson AJ. AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem . 2010;31(2):455–461. https://doi.org/10.1002/jcc.21334. Van-Wagenen BC, Larsen R, Cardellina JH, Randazzo D, Lidert ZC, Swithenbank C. Ulosantion, a potent insecticide from the sponge Ulosa ruetzleri. J Org Chem. 1993;58:335–337. https://doi.org/10.1021/jo00054a013. Viet TD, Xuan TD, Anh LH. α-Amyrin and β-Amyrin Isolated from Celastrus hindsii Leaves and Their Antioxidant, Anti-Xanthine Oxidase, and Anti-Tyrosinase Potentials. Molecules 2021;26:7248. https://doi.org/10.3390/ molecules26237248. World Health Organization. WHO guidelines on good agricultural and collection practices (GACP) for medicinal plants [internet]. Geneva; c2003 [cited 2023 Oct 16] Available from: https://iris.who.int/server/api/core/bitstreams/a00d3219-ed33-436a-be13-e3f00763f874/content Zhang P, Wang H, Xu X, Ye Y, Zhang Y. Correlation analysis between phytochemical composition and biological activities of Artemisia scoparia . Food Biosci. 2024;62:105342. https://doi.org/10.1016/j.fbio.2024.105342. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 14 Apr, 2026 Reviews received at journal 10 Apr, 2026 Reviews received at journal 10 Apr, 2026 Reviewers agreed at journal 03 Apr, 2026 Reviews received at journal 01 Apr, 2026 Reviewers agreed at journal 01 Apr, 2026 Reviewers agreed at journal 01 Apr, 2026 Reviewers agreed at journal 01 Apr, 2026 Reviewers agreed at journal 01 Apr, 2026 Reviews received at journal 05 Mar, 2026 Reviewers agreed at journal 03 Mar, 2026 Reviewers agreed at journal 02 Mar, 2026 Reviewers invited by journal 02 Mar, 2026 Editor invited by journal 17 Feb, 2026 Editor assigned by journal 17 Feb, 2026 Submission checks completed at journal 15 Feb, 2026 First submitted to journal 15 Feb, 2026 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. 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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-8786768","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":600632725,"identity":"c9703115-b8bc-415d-9754-6c283927b183","order_by":0,"name":"Uchenna Benjamin Okeke","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABFUlEQVRIiWNgGAWjYDACHgZmMG0AJisYEiBsNqK1nCFZC2MbEVr4eQ4fNvjAcFjenL332IOf8+zyDI4f3sDwoewwgzn/AqxaJHvbkhNnMBw23NlzLt2wd1tyscGZtALGGecOM1jOeIBVi8F5HuPDPAyHEwxu5JhJ8G5jTtxwIMeAmbftMIPBjQM4tPB/PvwHpOX+GzPJv3PqEzecf2PA/BeflrM9zMkMYFt4zKR5Gw4nbrgBtIURpOV8A3a/9BwzNuwxSDfccCbHTFrm2PFiyRvPCg4CPcZjcANXiCU/lvhRYS1vcPyMmeSbmuo8vvPJGx/8KLOWMziP3WFQ56HxQWp5GCQS8GjB4QJ8toyCUTAKRsEIAgBIcGRNMlNCGAAAAABJRU5ErkJggg==","orcid":"","institution":"Afe Babalola Universi","correspondingAuthor":true,"prefix":"","firstName":"Uchenna","middleName":"Benjamin","lastName":"Okeke","suffix":""},{"id":600632727,"identity":"f11b73e1-6c3e-4f03-8540-137cdfee8843","order_by":1,"name":"Patrick Igbinaduwa","email":"","orcid":"","institution":"University of Benin","correspondingAuthor":false,"prefix":"","firstName":"Patrick","middleName":"","lastName":"Igbinaduwa","suffix":""},{"id":600632729,"identity":"98388cc1-c148-4013-ae2a-d3870665151d","order_by":2,"name":"Ibrahim Oluwatobi Kehinde","email":"","orcid":"","institution":"University of KwaZulu-Natal","correspondingAuthor":false,"prefix":"","firstName":"Ibrahim","middleName":"Oluwatobi","lastName":"Kehinde","suffix":""},{"id":600632730,"identity":"1864e858-b92f-478b-95d1-5a21b33ab6c3","order_by":3,"name":"Vuyisa Mzozoyana","email":"","orcid":"","institution":"University of KwaZulu-Natal","correspondingAuthor":false,"prefix":"","firstName":"Vuyisa","middleName":"","lastName":"Mzozoyana","suffix":""}],"badges":[],"createdAt":"2026-02-04 12:55:10","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8786768/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8786768/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":104085128,"identity":"9303f296-27f8-4b37-bd20-804fb9e9b80d","added_by":"auto","created_at":"2026-03-06 15:11:50","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":196520,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eFicus sur\u003c/em\u003e in its natural habitat in Ibadan, Oyo, Nigeria\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/b114dda7224ab8df5af7bb1d.jpeg"},{"id":104085118,"identity":"1f196683-1ff0-4cf9-aee3-8094ef1f132d","added_by":"auto","created_at":"2026-03-06 15:11:47","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":250223,"visible":true,"origin":"","legend":"\u003cp\u003eFlow chart for the activity-guided separation, isolation, and purification of bioactive compounds from \u003cem\u003eFicus sur \u003c/em\u003ehydroethanol stem bark extract\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/78aaf35a63ae9d5e4547b30a.jpeg"},{"id":104085144,"identity":"f3f7aba3-31ff-41e2-8944-d7d7d6d38e72","added_by":"auto","created_at":"2026-03-06 15:11:54","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":130940,"visible":true,"origin":"","legend":"\u003cp\u003eCrystal structures of: (a) Steroid 5-alpha-reductase 2 (PDB ID: 7BW1) in complex with Finasteride; (b) \u003ca href=\"https://www.rcsb.org/structure/7YMJ\"\u003ealpha-1\u003csub\u003eA\u003c/sub\u003e adrenergic receptor R-Nb6 (PDB ID: 7YMJ) complex bound to tamsulosin\u003c/a\u003e; (c) Phosphodiesterase-5 (PDE5) (PDB ID: 2H42) in complex with sildenafil; and (d) the human M1 muscarinic acetylcholine receptor PDB ID: 5CXV) bound to antagonist tiotropium \u003ca href=\"https://www.rcsb.org/\"\u003e(https://www.rcsb.org/)\u003c/a\u003e.\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/1459d4f06419e1500f735800.jpeg"},{"id":104085120,"identity":"6a5df5f6-a056-4556-9837-1e0f52a53484","added_by":"auto","created_at":"2026-03-06 15:11:48","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":160646,"visible":true,"origin":"","legend":"\u003cp\u003e\u003csup\u003e1\u003c/sup\u003eH-NMR spectrum of compound \u003cstrong\u003eC5\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/967306eafb50bd47a8aa5457.png"},{"id":104085115,"identity":"3e458e86-00db-464c-baa0-81fc1200f971","added_by":"auto","created_at":"2026-03-06 15:11:46","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":225890,"visible":true,"origin":"","legend":"\u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC-NMR spectrum of \u003cstrong\u003eC5\u003c/strong\u003e (600 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ \u003csub\u003eppm\u003c/sub\u003e\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/7c01a7c414e6e1b626d0732b.png"},{"id":104085123,"identity":"c271cf2d-8e17-469d-b478-9b8f37ac5954","added_by":"auto","created_at":"2026-03-06 15:11:49","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":110919,"visible":true,"origin":"","legend":"\u003cp\u003eDEPTQ spectrum of compound \u003cstrong\u003eC5\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"floatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/badecec3ade77b8151164d89.png"},{"id":104085130,"identity":"93965516-5626-40f6-8862-e0cc3074a748","added_by":"auto","created_at":"2026-03-06 15:11:51","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":162840,"visible":true,"origin":"","legend":"\u003cp\u003e\u003csup\u003e1\u003c/sup\u003eH-NMR spectrum of \u003cstrong\u003eC8 \u003c/strong\u003e(600 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ \u003csub\u003eppm\u003c/sub\u003e\u003c/p\u003e","description":"","filename":"floatimage7.png","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/b9a53407f89641bb814cb393.png"},{"id":104085103,"identity":"10accc48-7764-4620-b11a-ac268ade6068","added_by":"auto","created_at":"2026-03-06 15:11:42","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":269098,"visible":true,"origin":"","legend":"\u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC-NMR spectrum of \u003cstrong\u003eC8\u003c/strong\u003e (600 MHz, CDCl\u003csub\u003e3\u003c/sub\u003e) δ \u003csub\u003eppm\u003c/sub\u003e\u003c/p\u003e","description":"","filename":"floatimage8.png","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/b78da9eaa4826db51afed1e5.png"},{"id":104085126,"identity":"0bcbee83-a0ee-410c-ba03-a2b5c0f7c236","added_by":"auto","created_at":"2026-03-06 15:11:50","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":77757,"visible":true,"origin":"","legend":"\u003cp\u003eDEPTQ spectrum of compound \u003cstrong\u003eC8\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"floatimage9.png","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/d62c1db176af1b4a36c27f0b.png"},{"id":104085119,"identity":"9840c0be-ad25-418c-a5ab-5b77aacbf9ba","added_by":"auto","created_at":"2026-03-06 15:11:47","extension":"jpeg","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":166783,"visible":true,"origin":"","legend":"\u003cp\u003eStructures of compound: (A) α-Amyrin acetate; (B) β-Sitosterol\u003c/p\u003e","description":"","filename":"floatimage10.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/46781ae3da04ae93d7147c80.jpeg"},{"id":104085114,"identity":"54eb944d-f10c-4d63-ac2d-1b22b63ff0c4","added_by":"auto","created_at":"2026-03-06 15:11:46","extension":"jpeg","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":95277,"visible":true,"origin":"","legend":"\u003cp\u003eCOSY spectrum of compound \u003cstrong\u003eC5 \u003c/strong\u003edisplaying \u003csup\u003e1\u003c/sup\u003eH - \u003csup\u003e1\u003c/sup\u003eH interactions\u003cstrong\u003e (\u003c/strong\u003eyellow circle: H-2 and H-3; red circle: H-11 and H-12).\u003c/p\u003e","description":"","filename":"floatimage11.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/89c2c94f9a067464389d2ea0.jpeg"},{"id":104085131,"identity":"0d840ba8-005e-444b-8a9b-1422dfd27e5e","added_by":"auto","created_at":"2026-03-06 15:11:51","extension":"jpeg","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":115437,"visible":true,"origin":"","legend":"\u003cp\u003eCOSY spectrum of compound \u003cstrong\u003eC8 \u003c/strong\u003edisplaying \u003csup\u003e1\u003c/sup\u003eH - \u003csup\u003e1\u003c/sup\u003eH interactions\u003cstrong\u003e \u003c/strong\u003e(Green circle: H-2 and H-3; Yellow circle: H-6 and H-7).\u003c/p\u003e","description":"","filename":"floatimage12.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/9e7f4e075071b4b3a8700347.jpeg"},{"id":104085122,"identity":"752cdb6d-61e8-49ac-95fc-baabf5870a2e","added_by":"auto","created_at":"2026-03-06 15:11:49","extension":"jpeg","order_by":13,"title":"Figure 13","display":"","copyAsset":false,"role":"figure","size":531308,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003e2D interaction map of α-amyrin acetate (A) complex with BPH target proteins - 5α-reductase type 2 (7BW1), α1A-adrenoceptor (7YMJ), phosphodiesterase-5 (2H42), and muscarinic M3 receptor (5ZHP).\u003c/em\u003e\u003c/p\u003e","description":"","filename":"floatimage13.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/0ba2d78aa4991a6db549a338.jpeg"},{"id":104085132,"identity":"3a1d090d-c960-4c98-ab39-8ce0979e6ba8","added_by":"auto","created_at":"2026-03-06 15:11:52","extension":"jpeg","order_by":14,"title":"Figure 14","display":"","copyAsset":false,"role":"figure","size":143079,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003e2D interaction map of β-sitosterol (B) complex with BPH target proteins - 5α-reductase type 2 (7BW1), α1A-adrenoceptor (7YMJ), phosphodiesterase-5 (2H42), and muscarinic M3 receptor (5ZHP).\u003c/em\u003e\u003c/p\u003e","description":"","filename":"floatimage14.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/979ff4d18dd99ad62c765658.jpeg"},{"id":104085125,"identity":"dd63ba8d-1df5-478f-95a2-558bda5516ee","added_by":"auto","created_at":"2026-03-06 15:11:49","extension":"jpeg","order_by":15,"title":"Figure 15","display":"","copyAsset":false,"role":"figure","size":268318,"visible":true,"origin":"","legend":"\u003cp\u003eADMETLab evaluation showing the suitable physicochemical space for oral bioavailability; A: α-amyrin acetate; B: β-sitosterol\u003c/p\u003e","description":"","filename":"floatimage15.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/19ff46c1b7e9f5f0debe2fe9.jpeg"},{"id":104402459,"identity":"c3bf71ea-20f3-403f-bb0c-8ea99a9563fc","added_by":"auto","created_at":"2026-03-11 12:15:26","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4132497,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8786768/v1/6692f845-fd84-402e-a952-b7a7468c4b3d.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Bioactive triterpenoids isolated and characterized from Ficus sur (Forrsk) stem bark extract demonstrate inhibitory activity against benign prostatic hyperplasia protein targets using in silico study","fulltext":[{"header":"1 Introduction","content":"\u003cp\u003eThe search for chemical entities from natural products for the management of emerging and complex disease conditions has continued to experience a quantum leap, creating a basis for more extensive scientific exploration of diverse lead structures that serve as templates for developing new pharmaceuticals. The growing interest in herbal medicines is largely driven by the perception that they are safer and more affordable than synthetic drugs, which are often expensive and associated with adverse side effects. Studies have revealed that various phytochemicals, such as alkaloids, flavonoids, phenolics, terpenoids, and saponins, found in medicinal plants can trigger a range of biological activities in living organisms, including antioxidant, anti-inflammatory, antibacterial, antiviral, and antitumor effects (Zhang et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). In vitro and in vivo studies validated these hypotheses that plant-derived compounds act through multiple pathways (Latif and Nawaz, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), including inhibition of 5α-reductase, a key enzyme in benign prostatic hyperplasia and prostate cancer pathogenesis, due to their structural similarity to endogenous receptor ligands (Nguyen et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eBenign prostatic hyperplasia (BPH), a nonmalignant enlargement of the prostate causing lower urinary tract symptoms (LUTS), has become a significant health issue among aging men worldwide (Speakman et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Due to the bothersome side effects linked to conventional medications, the demand for herbal treatments for BPH management has greatly increased. Herbal remedies, some available in formulated dosage forms, have demonstrated effectiveness in treating BPH, cancer chemotherapy, and chemoprophylaxis. For example, herbal preparations containing either single or combined extracts of \u003cem\u003eSaw palmetto, Serenoa repens, Pygeum africanum, Urtica dioica, Cucurbita pepo\u003c/em\u003e, and β-sitosterol from \u003cem\u003eHypoxis rooper\u003c/em\u003e (African star grass) help manage BPH (Csik\u0026oacute;s et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eBiological activity-guided isolation techniques have produced important therapeutic agents for diseases like cancer, Alzheimer\u0026rsquo;s disease, infections, malaria, and diabetes (Atanasov et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Advances in computer-aided drug design have further sped up the discovery and development of plant-derived drugs, making them increasingly promising. To achieve significant improvements and increase cost-efficiency in drug development, employing computational methods has become essential. Typically, after identifying and selecting disease-related targets, various computational tools perform key drug discovery steps, including target identification (such as an enzyme or protein receptor), modifying drug-like compounds, and optimizing these compounds to develop effective drug candidates (Duffy et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eFicus sur\u003c/em\u003e, also known as broom cluster fig or cape fig, belongs to the genus Moraceae and is widely distributed in several tropical African countries. The plant, due to its high medicinal richness, has been used in ethnomedicine to treat various disease conditions such as infections, tumors, pains, and urinary disorders (Sieniawska et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Several phytochemicals, such as triterpenoids, polyphenols, and fatty acids, have been isolated from different parts of \u003cem\u003eF.\u003c/em\u003e sur in preceding studies (Ogunlaja, 2022; Sieniawska et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Several phytosterol (β-sitosterol, stigmasterol), oleanane-type (12-oleanen-3-yl acetate, oleanen-3-yl acetate), and ursine-type (urs-12-en-24-oic acid, α-amyrin, 3-acetoxy-α-amyrin) triterpenoids have been isolated from the latex, leaf, and stem bark of \u003cem\u003eF. sur\u003c/em\u003e (Nwanisobi et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Ogunlaja et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Sieniawska et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Mouelle et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Okeke et al (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2026\u003c/span\u003e) reported the presence of gamma-sitosterol from the \u003cem\u003eF. sur\u003c/em\u003e hydroethanolic stem bark extract. However, there is a paucity of scientific validation of the efficacy of these phytochemicals on benign prostatic hyperplasia. This work reports herein the isolation and characterization of two triterpenoids, α-amyrin acetate and β-sitosterol, from the ethyl acetate fraction of the hydroethanolic stem bark of \u003cem\u003eF. sur\u003c/em\u003e and \u003cem\u003ein silico\u003c/em\u003e evaluation of their ameliorating activity against protein targets implicated in BPH pathogenesis.\u003c/p\u003e"},{"header":"2 Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Collection, extraction, and fractionation of plant material\u003c/h2\u003e \u003cp\u003e\u003cem\u003eF. sur\u003c/em\u003e stem bark was identified and collected from its natural habitat in Ibadan, Oyo State, Nigeria (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), by Dr. Akinniyi Samuel Odewo, a Taxonomist at the Forest Research Institute of Nigeria (FRIN), Ibadan, with compliance to WHO guidelines on good agricultural and collection practices for medicinal plants (World Health Organization, 2003). A herbarium specimen of the plant material (FHI113940) was deposited in the facility. The powdered stem bark (5 kg) was extracted by maceration with 80% ethanol at room temperature for 72 hours. The solution was filtered using Whatman No. 4 filter paper, and the filtrate was concentrated to dryness under vacuum at 50\u0026deg;C using a rotary evaporator to afford \u003cem\u003eF. sur\u003c/em\u003e hydroethanol stem bark extract (FSSE) (yield: 286 g, 5.72% \u003csup\u003ew\u003c/sup\u003e/\u003csub\u003ew\u003c/sub\u003e).\u003c/p\u003e \u003cp\u003eEmploying the modified Kupchan method of solvent partitioning as described by Van-Wagenen et al. (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e1993\u003c/span\u003e), 250 g of the extract initially suspended in 200 mL of distilled water was transferred completely into a 2000 mL separating funnel and successively partitioned in the order: \u003cem\u003en\u003c/em\u003e-hexane (HEX), dichloromethane (DCM), ethyl acetate (EtOAc), and \u003cem\u003en\u003c/em\u003e-butanol (BuOH). The various fractions collected were concentrated under vacuum in a rotary evaporator to obtain the concentrates, which were then air-dried and weighed.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Isolation and purification of compounds\u003c/h2\u003e \u003cp\u003eFirstly, the ethyl acetate fraction (40.00 g) of FSSE was adsorbed onto silica gel (20.00 g, 70\u0026ndash;230 mesh size) in a mortar to form a slurry, which was then air-dried and loaded into a 50 x 10 cm glass column packed with 500 g of silica gel (70\u0026ndash;230 mesh). Dichloromethane (100%) was used to pack the column before loading the sample. The column was eluted with gradient solvent systems of increasing polarity in the order: \u003cem\u003en\u003c/em\u003e-hexane, dichloromethane, and ethyl acetate. A total of 32 fractions were collected (250 mL each) until the compounds were completely eluted from the column. The fractions were monitored by thin-layer chromatography (TLC), and those with similar TLC profiles were pooled and reduced \u003cem\u003ein vacuo\u003c/em\u003e at 45\u0026deg;C to obtain 7 fractions (FSE-1 to FSE-7). Fractions FSE-1 (1\u0026ndash;5), FSE-2 (6\u0026ndash;9), and FSE-4 (13\u0026ndash;14) were subjected to a series of open column chromatography (CC) and preparative thin-layer chromatography (PTLC) using gradient solvent systems. The various sub-fractions obtained were monitored with TLC to confirm the presence of a single spot. The isolated compounds (C1 \u0026ndash; C8) (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) were identified by TLC and melting point determination. The structures of the compounds were elucidated through spectral analysis, including ultraviolet (UV), Fourier Transform Infrared (FTIR), \u0026sup1;H and \u0026sup1;\u0026sup3;C NMR, and DEPTQ spectroscopic experiments, and by comparing the spectral data with literature values.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Molecular docking of isolated compounds\u003c/h2\u003e \u003cp\u003eThe chemical structures of the isolated and characterized compounds were drawn using ChemDraw and converted to 3D structures. The compounds were optimized by correcting stereochemistry, removing duplicates, and adjusting protonation states to physiological pH (Omotuyi et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Molecular docking was performed using AutoDock Vina (version 1.2.3) to predict the binding affinities and interaction modes of the compounds with the BPH-related protein targets: 5α-reductase 2 (5αR2; PDB ID: 7BW1), α\u003csub\u003e1A\u003c/sub\u003e-adrenoceptor (ADRA1A; PDB ID: 7YMJ), phosphodiesterase-5 (PDE5; PDB ID: 2H42), and muscarinic M3 receptor (PDB ID: 5ZHP) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e) (Eberhardt et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Trott and Olson, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Before docking, the prepared protein and ligand structures were converted into the PDBQT format using AutoDock Tools (ADT, version 1.5.7). Docking calculations were carried out for each protein target using the prepared ligands within the defined grid box. For each ligand, AutoDock Vina generated multiple binding poses ranked by predicted binding affinity (kcal/mol). Docking results were visualized and analyzed for protein\u0026ndash;ligand interactions using UCSF Chimera and BIOVIA Discovery Studio Visualizer (Pettersen \u003cem\u003eet al.\u003c/em\u003e, 2004).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Drug-likeness and \u003cem\u003ein-silico\u003c/em\u003e ADMET prediction analysis of isolated compounds\u003c/h2\u003e \u003cp\u003eADMETlab 3.0 Web tool (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://admetlab3.scbdd.com/\u003c/span\u003e\u003cspan address=\"https://admetlab3.scbdd.com/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) was used to predict the physicochemical properties, toxicity profile, and drug-likeness nature of the isolated compounds. The drug-likeness was evaluated based on the Lipinski rule, which is related to the compounds' predicted absorption, distribution, metabolism, and excretion (ADME) properties. Compounds with multiple violations are generally less likely to demonstrate favourable \u003cem\u003ein vivo\u003c/em\u003e activity. Topological polar surface area (TPSA), another critical pharmacokinetic parameter, was also examined, as it predicts membrane permeability of polar atoms; optimal passive absorption typically falls within the range of 20\u0026ndash;140 \u0026Aring;\u0026sup2; (Fu et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e"},{"header":"3 Results and Discussion","content":"\u003cp\u003eOpen column chromatography (CC) and PTLC separation methods on Silica gel afforded the isolation of majorly semi-polar constituents (C1 to C8) from the ethyl acetate fraction of FSSE. The structure of the isolated compounds was confirmed using various spectral analysis techniques like \u0026sup1;H NMR, \u0026sup1;\u0026sup3;C NMR, DEPTQ, and comparison of the spectra data with those in the literature.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Physical characteristics and identification of isolated compounds\u003c/h2\u003e \u003cp\u003e \u003cb\u003eCompound C5\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cem\u003eα-Amyrin acetate.\u003c/em\u003e Isolated and purified as a 240 mg white solid, soluble in 99% chloroform, with a melting point of 225\u0026ndash;227\u0026deg;C and a positive Liebermann-Buchard test for steroids.\u003c/p\u003e \u003cp\u003e \u003cstrong\u003eSpectral Analysis\u003c/strong\u003e \u003cp\u003eUV-vis spectrum (CHCl\u003csub\u003e3\u003c/sub\u003e solvent) recorded λmax at 227 nm in chloroform. The FTIR spectrum showed absorption bands (in cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) at 2952.1, 2911.1, 1733.2, 1244.9, and 1457.4, respectively. This reflects the presence of key functional groups in the compound. Pansare et al. (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) reported a similar FTIR absorbance pattern for α-amyrin acetate.\u003c/p\u003e \u003c/p\u003e \u003cp\u003eThe \u003csup\u003e1\u003c/sup\u003eH NMR spectrum (CDCl\u003csub\u003e3\u003c/sub\u003e solvent; 600 MHz NMR spectrometer) of C5 shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e, revealed various proton peaks as follows (δ in ppm): 5.18 (1H, \u003cem\u003et\u003c/em\u003e, H-12), 4.51 (1H, \u003cem\u003et\u003c/em\u003e, H-3), 2.30 (3H, \u003cem\u003es\u003c/em\u003e, H-2'), 2.01 (1H, \u003cem\u003em\u003c/em\u003e, H-20), 1.90 (2H, \u003cem\u003et\u003c/em\u003e, H-11), 1.63 (2H, \u003cem\u003em\u003c/em\u003e, H-2), 1. 61 (1H, \u003cem\u003et\u003c/em\u003e, H-9), 1.53 (2H, \u003cem\u003et\u003c/em\u003e, H-6). Notably, the deshielded vinyl protons H-12 and H-3 peaks appeared downfield as a triplet at 5.15 ppm and a doublet of doublets at 4.50, respectively. A sharp, intense singlet at 2.10 ppm corresponds to the three methyl protons of an acetate group, confirming the presence of an acetate moiety. These spectral features are consistent with an ursane-type pentacyclic triterpenoid, in contrast to the oleanane-type (e.g., β-amyrin), with a H-12 proton resonating near 5.18 ppm. Eight methyl signals are clustered in a crowded upfield region between 0.65 and 1.20.\u003c/p\u003e \u003cp\u003eThe \u003csup\u003e13\u003c/sup\u003eC NMR and DEPTQ spectra in Figs.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e and \u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e revealed a total of 32 carbon signals, categorized as eight methyl (-CH\u003csub\u003e3\u003c/sub\u003e), nine methylene (-CH\u003csub\u003e2\u003c/sub\u003e), seven methine (-CH), and seven quaternary (\u0026ndash;C-) carbons, detailed as follows (CDCl\u003csub\u003e3\u003c/sub\u003e solvent; 600 MHz NMR spectrometer): δ ppm 170.95 (C-1'), 139.64 (C-13), 124.34 (C-12), 80.97 (C-3), 59.10 (C-18), 55.30 (C-5), 47.67 (C-9), 42.10 (C-14), 41.74 (C-8), 40.06 (C-22), 39.83 (C-1). The chemical shift values presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e align closely with those reported for α-amyrin acetate by Okoye et al. (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2014\u003c/span\u003e), indicating concordance between the observed and literature NMR spectra. A characteristic signal at 170.92 ppm indicates a carbonyl carbon (C\u0026thinsp;=\u0026thinsp;O) of an ester. Peaks observed downfield at 124.34 and 139.64 ppm correspond to the olefinic C-12 methine and C-13 quaternary carbons, respectively. The methine carbons C-18 and C-19 appeared downfield at 59.10 and 39.84 ppm. These assignments further support the characterization of the compound as an ursane-type triterpenoid, distinguishing it from the oleanane framework, which typically shows C-12 and C-13 resonance peaks near 121 and 145 ppm, and C-18 and C-19 signals nearly overlapping around 47\u0026ndash;47.5 ppm (Okoye et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Viet et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). \u003csup\u003e1\u003c/sup\u003eH-\u003csup\u003e1\u003c/sup\u003eH COSY of C5 (Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e11\u003c/span\u003e) revealed cross-peaks indicating coupling between neighboring protons, particularly H-2 / H-3 (1.6 and 4.55 ppm), H-11 / H-12 (1.9 and 5.15 ppm), and H-12 / H-18 (5.15 and 1.2 ppm).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC-NMR and \u003csup\u003e1\u003c/sup\u003eH-NMR data of \u003cb\u003eC5\u003c/b\u003e and α-amyrin acetate from literature\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePosition\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eChemical shift, δ (ppm) value\u003c/p\u003e \u003cp\u003eCompound C5 (δ ppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eChemical shift, δ (ppm) value\u003c/p\u003e \u003cp\u003eCompound C5 (δ ppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eα-amyrin acetate (Okoye et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2014\u003c/span\u003e)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003csup\u003e1\u003c/sup\u003eH\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003csup\u003e1\u003c/sup\u003eH\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC NMR\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.63 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.55 \u003cem\u003e(dd)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e80.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.56 \u003cem\u003e(dd)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e80.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e81.18\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e55.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e55.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e55.46\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.53 \u003cem\u003e(t)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e39.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e39.83\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.61 \u003cem\u003e(t)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e47.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e47.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e47.84\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.90 \u003cem\u003e(t)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.15 \u003cem\u003e(t)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e124.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.18 \u003cem\u003e(t)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e124.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e124.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e139.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e139.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e139.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e42.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e42.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e42.10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e59.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e59.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e59.26\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e39.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.01 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1'\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e170.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e170.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e171.53\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2'\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eChemical shift values expressed as parts per million (ppm); \u003cem\u003es-\u003c/em\u003e singlet, \u003cem\u003ed-\u003c/em\u003e doublet, \u003cem\u003et-\u003c/em\u003e triplet, \u003cem\u003edd-\u003c/em\u003e doublet of a doublet, \u003cem\u003em-\u003c/em\u003e multiplet\u003c/p\u003e \u003cp\u003e \u003cb\u003eCompound C8\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cem\u003eβ-Sitosterol.\u003c/em\u003e Isolated and purified as a 341 mg pale yellow crystalline solid, soluble in 99% chloroform, with a melting point of 136\u0026ndash;138\u0026deg;C, and a positive Liebermann-Buchard test for steroids.\u003c/p\u003e \u003cp\u003e \u003cem\u003eSpectral analysis\u003c/em\u003e: UV-vis spectrum (CHCl\u003csub\u003e3\u003c/sub\u003e solvent) recorded λmax at 260 nm, principally attributed to a single isolated double bond and the solvent effect (bathochromic shift) of chloroform. \u003csup\u003e1\u003c/sup\u003eH NMR spectrum (CDCl\u003csub\u003e3\u003c/sub\u003e solvent, 600 MHz NMR) revealed various proton peaks at different chemical shifts (δ in ppm) as follows: 5.3 (1H, \u003cem\u003es\u003c/em\u003e, H-6), 3.48 (1H, \u003cem\u003em\u003c/em\u003e, H-3), 2.2 (2H, \u003cem\u003em\u003c/em\u003e, H-2), 2.2 (2H, \u003cem\u003em\u003c/em\u003e, H-4), 1.9 (2H, \u003cem\u003et\u003c/em\u003e, H-7) (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e). Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e reveals chemical shift values that correspond closely with the β-sitosterol NMR spectrum documented by Ododo et al. (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) and Javed et al. (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), confirming NMR spectral consistency between the observed and literature values. A distinct multiplet peak at 5.36 ppm corresponds to the olefinic proton on carbon C-6. The deshielded hydroxyl group proton on carbon C-3 appears as a doublet of doublets at 3.49 ppm.\u003c/p\u003e \u003cp\u003e \u003csup\u003e13\u003c/sup\u003eC NMR and DEPTQ spectra of compound C5 in Figs.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e and \u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e9\u003c/span\u003e, revealed a total of 29 carbon signals as six methyl (-CH\u003csub\u003e3\u003c/sub\u003e), eleven methylene (-CH\u003csub\u003e2\u003c/sub\u003e), nine methine (-CH) and three quaternary (-C-) carbons, detailed as follows (600 MHz, in CDCl\u003csub\u003e3\u003c/sub\u003e solvent) δ ppm: 140.76 (C-5), 129.30 (C-6), 71.81 (C-3), 56.89 (C-17), 56.79 (C-14), 50.1 (C-9), 45.87 (C-24), 42.33 (C-13), 42.3 (C-4), 40.47 (C-12), 37.20 (C-1), 36.51(C-10), 36.15 (C-20). \u003csup\u003e1\u003c/sup\u003eH-\u003csup\u003e1\u003c/sup\u003eH COSY revealed cross-peaks indicating correlations between neighboring protons in compound C8 (Fig.\u0026nbsp;\u003cspan refid=\"Fig12\" class=\"InternalRef\"\u003e12\u003c/span\u003e), particularly H-2 / H-3 (2.3 and 3.49 ppm), H-3 / H-4 (3.39 ppm and 2.29 ppm), and H-6 / H-7 (5.35 and 1.98 ppm).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC-NMR and \u003csup\u003e1\u003c/sup\u003eH-NMR data of \u003cb\u003eC8\u003c/b\u003e and β-sitosterol from literature\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePosition\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eCompound C8 chemical shift, δ (ppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eβ-Sitosterol\u003c/p\u003e \u003cp\u003e(Javed et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2021\u003c/span\u003e )\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eβ-Sitosterol\u003c/p\u003e \u003cp\u003e(Ododo et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2016\u003c/span\u003e )\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eβ-Sitosterol\u003c/p\u003e \u003cp\u003e(Javed et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2021\u003c/span\u003e )\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eβ-Sitosterol\u003c/p\u003e \u003cp\u003e(Ododo et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2016\u003c/span\u003e )\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003csup\u003e1\u003c/sup\u003eH\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003csup\u003e1\u003c/sup\u003eH NMR\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003csup\u003e1\u003c/sup\u003eH NMR\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC NMR\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003csup\u003e13\u003c/sup\u003eC NMR\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e37.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.46 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e36.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e37.28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.3 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.56 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e31.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e31.69\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.49 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e71.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.54 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e72.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e71.82\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.29 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e42.23 \u003cem\u003e(t)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.32 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e42.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e42.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e140.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e141.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e140.77\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.35 \u003cem\u003e(t)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e121.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.37 \u003cem\u003e(t)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e122.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e121.73\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.98 \u003cem\u003e(t)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e31.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.04 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e31.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e31.93\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e31.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.69 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e31.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e31.93\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e50.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.55 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e51.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50.16\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e36.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e35.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e36.51\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.52 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colname=\"c1\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e31.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.93 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e32.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e33.98\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e26.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.15 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e25.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e26.11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e45.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.38 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e46.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e45.86\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e29.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.57 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e30.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e29.19\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.84 \u003cem\u003e(d)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e21.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e19.84\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.86 \u003cem\u003e(d)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e19.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e19.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.10 \u003cem\u003e(m)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e23.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e23.10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.82 \u003cem\u003e(t)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e12.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e12.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC-3 OH\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.98 \u003cem\u003e(s)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eChemical shift values expressed as parts per million (ppm); \u003cem\u003es-\u003c/em\u003e singlet, \u003cem\u003ed-\u003c/em\u003e doublet, \u003cem\u003et-\u003c/em\u003e triplet, \u003cem\u003em-\u003c/em\u003e multiplet\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Molecular docking analysis of isolated bioactive compounds against BPH targets\u003c/h2\u003e \u003cp\u003eThe docking scores, representing the predicted binding free energies (kcal/mol) of α-amyrin acetate and β-sitosterol, isolated from the ethyl acetate fraction of FSSE, against key therapeutic targets implicated in benign prostatic hyperplasia (BPH), are presented in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMolecular docking scores (kcal/mol) of α-amyrin acetate and β-sitosterol against BPH targets\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCompounds\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5α reductase 2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eα\u003csub\u003e1A\u003c/sub\u003e- adrenoceptor\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePhosphodiesterase-5\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMuscarinic M3\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePDB ID: 7BW1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePDB ID: 7YMJ\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePDB ID: 2H42\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePDB ID: 5ZHP\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eα-Amyrin acetate (C5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e-10.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e-10.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e-9.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e-8.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eβ-Sitosterol (C8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e-10.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e-8.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e-6.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e-7.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003ePDB \u0026ndash; Protein Data Bank\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Molecular interaction analysis of \u003cem\u003eFicus sur\u003c/em\u003e bioactive compounds with BPH target proteins\u003c/h2\u003e \u003cp\u003eFigure\u0026nbsp;\u003cspan refid=\"Fig13\" class=\"InternalRef\"\u003e13\u003c/span\u003e and \u003cspan refid=\"Fig14\" class=\"InternalRef\"\u003e14\u003c/span\u003e depicted the two-dimensional (2D) interaction profiles of the isolated compounds in Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e10\u003c/span\u003e, docked against the four major therapeutic targets of BPH. The visualized interactions highlight a combination of Van der Waals forces, π\u0026ndash;π stacking, π\u0026ndash;alkyl, alkyl, and hydrogen bonding, which collectively contribute to the stability and specificity of the ligand\u0026ndash;receptor complexes, corroborating the docking energy results in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e3\u003c/span\u003e. α-Amyrin acetate complexes displayed dense networks of hydrophobic and aromatic interactions across all receptor pockets, indicating a strong binding affinity and multi-target potential. Within the 5α-reductase 2 complex (7BW1), α-amyrin acetate established extensive π\u0026ndash;alkyl and alkyl contacts with residues such as Phe118, Arg114, Leu23, and Val24, while also forming Van der Waals interactions with Ala26 and Gly115, stabilizing the ligand within the hydrophobic catalytic channel of the enzyme. In the α\u003csub\u003e1A\u003c/sub\u003e-adrenoceptor complex (7YMJ), the compound interacted prominently with Trp102, Tyr176, and Phe312 through π\u0026ndash;π and hydrophobic contacts, suggesting a strong occupation of the receptor\u0026rsquo;s transmembrane binding site that could contribute to adrenergic antagonism. Similarly, in the phosphodiesterase-5 (2H42) complex, α-amyrin acetate formed stabilizing interactions with Phe820, Met816, and Ala823, complemented by π\u0026ndash;alkyl contacts that mimic known inhibitor binding patterns in the cGMP catalytic domain. For the muscarinic M3 receptor (5ZHP), α-amyrin acetate maintained π\u0026ndash;π and Van der Waals interactions with residues Phe437, Arg109, and Asn107, suggesting that it may also influence bladder contractility through muscarinic receptor modulation. These interactions explain the consistently high docking energies observed (\u0026minus;\u0026thinsp;10.9 to \u0026minus;\u0026thinsp;8.0 kcal/mol) and reinforce the compound\u0026rsquo;s potential as a multi-target modulator in BPH.\u003c/p\u003e \u003cp\u003eβ-Sitosterol also exhibited favorable interaction patterns with all four BPH targets. In the 5α-reductase binding pocket, β-sitosterol engaged in π\u0026ndash;π stacking and alkyl interactions with key hydrophobic residues, including Phe118, Tyr176, and Leu23, which are crucial for enzyme inhibition and suppression of dihydrotestosterone biosynthesis. The α\u003csub\u003e1A\u003c/sub\u003e-adrenoceptor complex revealed π\u0026ndash;alkyl interactions with Phe312 and Trp102, along with hydrogen bonding to Tyr196, which collectively suggest strong antagonistic behavior similar to known α1-blockers. Within the phosphodiesterase-5 complex, β-sitosterol interacted mainly with hydrophobic residues Phe820, Met816, and Val826, while the M3 receptor complex exhibited π\u0026ndash;π stacking with Phe437 and hydrophobic contacts with Ile424 and Val426. The abundance of hydrophobic and aromatic contacts indicates that β-sitosterol\u0026rsquo;s lipophilic nature allows it to efficiently stabilize within receptor pockets, consistent with its recognized pharmacological role as a 5α-reductase inhibitor and prostate health modulator.\u003c/p\u003e \u003cp\u003eThe molecular interaction analysis confirms that the investigated phytoconstituents, α-amyrin acetate and β-sitosterol, exhibit strong and complementary binding profiles across multiple BPH-related targets. Their interactions are dominated by π\u0026ndash;π and hydrophobic forces involving catalytically and structurally relevant residues, which not only stabilize the complexes but also suggest a synergistic multi-target therapeutic mechanism. This molecular evidence supports the hypothesis that the bioactive constituents of \u003cem\u003eFicus sur\u003c/em\u003e exert anti-BPH activity through concurrent inhibition of androgenic (5α-reductase), adrenergic (α\u003csub\u003e1A\u0026minus;\u003c/sub\u003eadrenoceptor), phosphodiesterase (PDE-5), and muscarinic receptor pathways, offering a mechanistic rationale for the traditional use of \u003cem\u003eFicus sur\u003c/em\u003e in prostate and urinary-related disorders.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.4 Drug-Likeness and ADMET Properties of Isolated Compounds\u003c/h2\u003e \u003cp\u003eThe drug-likeness and pharmacokinetic profile of α-amyrin acetate (C5) and β-sitosterol (C8) isolated from ethyl acetate fraction of FSSE were evaluated using the ADMETlab 3.0 web interface, with reference to Lipinski\u0026rsquo;s rule of five (molecular mass (\u0026lt;\u0026thinsp;500 Da), lipophilicity (Log\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;5), hydrogen bond donors (HBD\u0026thinsp;\u0026le;\u0026thinsp;5), and hydrogen bond acceptors (HBA\u0026thinsp;\u0026le;\u0026thinsp;10) (Lipinski et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2001\u003c/span\u003e), along with additional pharmacokinetic descriptors. Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e3\u003c/span\u003e summarises the key parameters supporting the suitability of the isolated compounds for biological evaluation, including their predicted physicochemical properties, lipophilicity, water solubility, pharmacokinetics, and overall drug-likeness. α-Amyrin acetate and β-sitosterol demonstrated favourable oral bioavailability, obeying Lipinski\u0026rsquo;s rule of 5. The ADMETLab output for the two compounds showed a coloured zone that represents the suitable physicochemical space for oral bioavailability, based on key physicochemical descriptors. Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e3\u003c/span\u003e presents the critical parameters supporting Compounds C5 and C8 as candidates for biological evaluation. These features further suggest that both compounds are likely to demonstrate good oral bioavailability, which reinforces the potential of the compounds as therapeutic candidates for the treatment of BPH and other bladder disorders.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eADMET physicochemical properties of ligands with the best binding affinities\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProperty\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMeaning\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOptimal Value\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eα-Amyrin acetate\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eβ-Sitosterol\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eMW\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMolecular weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e150 mg/mol ˂ MW\u0026thinsp;\u0026le;\u0026thinsp;500 g/mol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e468.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e414.39\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLogP\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLipophilicity\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u0026ndash;3 log mol/L\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e*5.799\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e*8.004\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLogS\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAqueous solubility\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-4\u0026ndash;0.5 log mol/L\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-6.519\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-7.221\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLogD\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDistribution coefficient (pH 7.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1\u0026ndash;3 log mol/L (pH 7.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.211\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.37\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTPSA\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTopological polar surface area\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u0026thinsp;~\u0026thinsp;140A\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e26.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20.23\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003enHA\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber of hydrogen bond acceptors\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u0026thinsp;~\u0026thinsp;12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003enHD\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber of hydrogen bond donors\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u0026thinsp;~\u0026thinsp;7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003enRot\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber of rotatable bonds\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u0026thinsp;~\u0026thinsp;11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003enRing\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber of rings\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u0026thinsp;~\u0026thinsp;6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003enRig\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber of rigid bonds\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u0026thinsp;~\u0026thinsp;30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e27.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eMaxRing\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber of atoms in the biggest ring\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u0026ndash;18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e22.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e17.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003efCHar\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFormal charge\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-4\u0026thinsp;~\u0026thinsp;4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003enHet\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber of heteroatoms\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1\u0026thinsp;~\u0026thinsp;15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNLRV\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber of Lipinski\u0026rsquo;s violations\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003e* - \u003cb\u003eLRV\u003c/b\u003e: Lipinski\u0026rsquo;s rule violation; \u003cb\u003eNLRV\u003c/b\u003e: Number of Lipinski\u0026rsquo;s rule violations\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4 Conclusion","content":"\u003cp\u003eOpen column chromatography, preparative thin-layer chromatography, and instrumental analytical techniques such as UV, FTIR, \u003csup\u003e1\u003c/sup\u003eH and \u003csup\u003e13\u003c/sup\u003eC NMR, resulted in the isolation of an ursane-type pentacyclic triterpenoid, α-Amyrin acetate (C5), and a phytosterol, β-sitosterol (C8). Both compounds (α-amyrin acetate and β-sitosterol) exhibited ameliorative effects against BPH through molecular docking on BPH protein targets, with β-sitosterol displaying binding scores comparable to those of the standard BPH drug, finasteride. This study highlights the diversity of this plant, validation of its traditional use and potential as a source of lead compounds for developing therapeutics to treat BPH and other urinary-related conditions, and contribution to improvement in public health strategies for managing BPH, especially in developing regions where access to standard therapies may be limited.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors acknowledge the contributions of the Faculty of Pharmacy, University of Benin, Nigeria, and Afe Babalola University, Ado-Ekiti, Nigeria, for providing the facilities for conducting this research, and also the School of Chemistry and Physics, University of KwaZulu-Natal, Durban, South Africa, for providing the facilities for the NMR experiments.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eUchenna Benjamin Okeke: Research concept development, data collection, analysis, manuscript draft preparation, review, and editing. Patrick Igbinaduwa: Research concept development, data analysis, and critical reading of manuscript. Vuyisa Mzozoyana: NMR analysis and data collection. Ibrahim Oluwatobi Kehinde: Bioinformatics analysis, article draft review, and editing.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eThe authors declare that no funds, grants, or other support were received during the research work and preparation of this manuscript.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that the data supporting the findings of this study are available within the paper. Should any raw data files be needed in another format, the corresponding author, upon reasonable request and with the consent of the other authors of this research, shall make them available.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest. We confirm the publication of this article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics and consent to participate\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePermissions or licences for plant collection and animal use are not applicable in this study, as the plant is common and non-endangered. \u003cem\u003eFicus sur\u003c/em\u003e herbarium specimen (Voucher number: FHI113940) was deposited at Forest Herbarium Ibadan (FHI), located in the Taxonomy Section of the Department of Forest Conservation and Protection, Forestry Research Institute of Nigeria (FRIN), Ibadan. The plant identifier and collector at FRIN is Dr. Akinniyi Samuel Odewo.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAtanasov AG, Zotchev SB, Dirsch VM, Supuran CT. 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WHO guidelines on good agricultural and collection practices (GACP) for medicinal plants [internet]. Geneva; c2003 [cited 2023 Oct 16] Available from: https://iris.who.int/server/api/core/bitstreams/a00d3219-ed33-436a-be13-e3f00763f874/content\u003c/li\u003e\n\u003cli\u003eZhang P, Wang H, Xu X, Ye Y, Zhang Y. Correlation analysis between phytochemical composition and biological activities of \u003cem\u003eArtemisia scoparia\u003c/em\u003e. \u003cem\u003eFood Biosci. \u003c/em\u003e2024;62:105342. https://doi.org/10.1016/j.fbio.2024.105342.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"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":"discover-chemistry","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Discover Chemistry](https://link.springer.com/journal/44371)","snPcode":"44371","submissionUrl":"https://submission.nature.com/new-submission/44371/3","title":"Discover Chemistry","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Ficus sur, amyrin acetate, sitosterol, prostatic hyperplasia, triterpenoid","lastPublishedDoi":"10.21203/rs.3.rs-8786768/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8786768/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003ePlants possess an abundant reservoir of compounds that can be harnessed and repurposed as therapeutic agents for the management of existing and emerging disease conditions, such as benign prostatic hyperplasia (BPH). This study aimed to isolate and characterize bioactive phytochemicals from the ethyl acetate fraction of \u003cem\u003ethe Ficus sur\u003c/em\u003e hydroethanol stem bark extract (FSSE) and to carry out \u003cem\u003ein silico\u003c/em\u003e evaluation of their activity against key protein targets implicated in BPH pathogenesis. \u003cem\u003eFicus sur\u003c/em\u003e stem bark was collected from its natural habitat, authenticated, extracted, and partitioned into different solvent fractions (dichloromethane, ethyl acetate, and \u003cem\u003en-\u003c/em\u003ebutanol). Sequential open column chromatography and preparatory thin layer chromatography of the ethyl acetate fraction led to the isolation and purification of compounds, which were characterized by spectroscopic analysis (UV, FTIR, \u003csup\u003e1\u003c/sup\u003eH and \u003csup\u003e13\u003c/sup\u003eC-NMR, and DEPTQ experiments). An \u003cem\u003eIn-silico\u003c/em\u003e molecular docking study was conducted using Autodock Vina, while ADMETlab 3.0 was employed to predict the physicochemical properties and drug-likeness of the isolated compounds. Two triterpenoids, α-amyrin acetate (110 mg) and β-sitosterol (340 mg), from the eight isolated compounds were characterized. \u003cem\u003eIn silico\u003c/em\u003e analyses indicated that both compounds exhibited binding affinities and physicochemical profiles comparable to those of the co-crystallized ligand finasteride across the four molecular targets implicated in benign prostatic hyperplasia. This gave credence to the medicinal richness of this plant and its use in folklore for urinary-related disorders such as BPH. It has been established in this study that \u003cem\u003eF. sur\u003c/em\u003e stem bark extract is rich in bioactive compounds that can be explored for the discovery of leads for developing new drug candidates, especially for BPH management.\u003c/p\u003e","manuscriptTitle":"Bioactive triterpenoids isolated and characterized from Ficus sur (Forrsk) stem bark extract demonstrate inhibitory activity against benign prostatic hyperplasia protein targets using in silico study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-06 15:10:11","doi":"10.21203/rs.3.rs-8786768/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-04-14T09:05:53+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-10T07:54:20+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-10T04:31:59+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"244831051576909304423022049123614552179","date":"2026-04-03T11:39:33+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-01T17:11:34+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"16617374767473844246396186072216657448","date":"2026-04-01T13:38:33+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"330238172818160064190439152855743076812","date":"2026-04-01T09:30:24+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"59844073229310008370241284923280965531","date":"2026-04-01T08:01:47+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"102149480476698538950811624420180959770","date":"2026-04-01T07:30:13+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-03-05T20:47:34+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"46279958842658130171892245635423217495","date":"2026-03-03T12:38:22+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"86019126074158914933129945608166993319","date":"2026-03-02T16:40:29+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-03-02T10:13:24+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2026-02-17T13:14:45+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-17T10:28:12+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-02-15T23:32:14+00:00","index":"","fulltext":""},{"type":"submitted","content":"Discover Chemistry","date":"2026-02-15T23:28:01+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"discover-chemistry","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Discover Chemistry](https://link.springer.com/journal/44371)","snPcode":"44371","submissionUrl":"https://submission.nature.com/new-submission/44371/3","title":"Discover Chemistry","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a3bc0121-68d4-487f-8b4f-a1942920336e","owner":[],"postedDate":"March 6th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[],"tags":[],"updatedAt":"2026-05-19T09:25:05+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-06 15:10:11","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8786768","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8786768","identity":"rs-8786768","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}