{"paper_id":"0d823e91-c86b-4642-aec8-5c804dffc623","body_text":"From In Silico Design to Bioactivity: Synthesis and Pharmacological Profiling of 3-Substituted Quinazoline-2,4-dione Heterocycles | 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 From In Silico Design to Bioactivity: Synthesis and Pharmacological Profiling of 3-Substituted Quinazoline-2,4-dione Heterocycles Neelam Sharma, Nishtha Shalmali, Manish Kumar, Vijay Kumar Sharma, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8566641/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract A series of novel 3-substituted quinazoline-2,4(1H,3H)-dione derivatives was designed, synthesized, and evaluated for their anticonvulsant, antibacterial, and antifungal activities. Anticonvulsant efficacy was assessed in vivo using the maximal electroshock (MES) seizure model, while antibacterial and antifungal activities were screened against selected Gram-positive, Gram-negative bacterial, and fungal strains. To support the experimental findings and elucidate possible binding interactions, molecular docking studies were carried out against relevant biological targets for antibacterial (PDB ID: 6BPP), antifungal (PDB ID: 7VPR), and anticonvulsant activity (PDB ID: 2Z5X). Among the synthesized compounds, compound 4h exhibited significant anticonvulsant activity by effectively inhibiting extensor seizures in the MES model at a dose of 30 mg/kg, showing comparable efficacy to the reference drug phenytoin sodium. Compound 4f demonstrated broad-spectrum antibacterial activity against Gram-positive bacteria ( Bacillus subtilis , Staphylococcus aureus ) and Gram-negative bacteria ( Escherichia coli , Pseudomonas aeruginosa ), inhibiting microbial growth at 100 µg/mL. Additionally, compound 4b displayed potent antifungal activity against Candida albicans and Aspergillus niger at 100 µg/mL, surpassing the activity of the standard antifungal agent griseofulvin. The study highlights the quinazoline-2,4-dione heterocyclic scaffold as a promising pharmacophore for the development of multifunctional bioactive agents with potential therapeutic relevance and reduced adverse effects. The combined in silico and experimental findings provide a valuable foundation for further structural optimization and pharmacological exploration. Quinazoline-2 4-dione Heterocyclic scaffold Molecular docking Anticonvulsant activity Antimicrobial agents Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 1. Introduction Quinazolinedione is a heterocyclic moiety present in many naturally occurring alkaloids isolated from plant species like Zanthoxylum arborescens [ 1 ], Evodia officinalis [ 2 ] and from Phylloperthadiversa (chafer beetle) [ 3 ]. Many synthetic drugs like ketanserin (anti-hypertensive) [ 1 , 4 ], benzouracil (anti-viral) [ 5 ], selurampanel (anti-convulsant) [ 6 ], etc. are stated to contain quinazolinedione moiety (Fig. 1 ). Moreover, quinazolinedione derivatives reported for activities like anti-inflammatory [ 7 ], anti-serotonergic [ 8 ], anti-microbial [ 9 ], anti-cancer [ 10 , 11 ] etc. It is evident that quinazolinediones act as anticonvulsant by competitively binding to glutamate receptor, kainate receptor and AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid) receptor. This leads to reduction in glutamate binding affinity for the receptors and sodium ion channels remain closed and thus inhibition of seizures [ 12 ]. The literature survey revealed that quinazolinedione moiety if present in any compound shows anticonvulsant action due to following features mentioned in Fig. 2 [ 13 – 16 ]: hydrophobic aryl domain (Ar), hydrogen bonding domain (HBD) in the form of –NHCO- grouping and electron donor atom (D). Based on these features, a series of 3-substitutedquinazoline-2,4(1H,3H)-dione analogues were synthesized and screened for anti-convulsant activity against maximal electroshock (MES) convulsions. These derivatives were also screened for anti-bacterial and anti-fungal activity against selected bacterial and fungal strains. 2. Material and Methods 2.1. Chemistry Isatoic anhydride and triphosgene were acquired from Sigma-Aldrich (India), while remaining reagents were sourced from Merck which is located in India and Central Drug House (Delhi, India). A digital melting point device was utilised in order to ascertain the melting points of each and every compound that was synthesized. Silica Gel GF 256 (Merck) was used to coat TLC plates and TLC spots of synthesized compounds were visualized using TLC UV Cabinet (HMG, India). The 1 HNMR and 13 CNMR spectra were recorded on a Bruker Advance II NMR (400 MHz) Spectrometer in DMSO-d 6 at Punjab University, Chandigarh, Punjab, India. IR spectra were recorded on a Perkin Elmer FTIR 1650 Spectrometer. Mass spectra were recorded on Bruker MALDI Ultra Flextreme using 2,5-dihydroxybenzoic acid (DHB) matrix at Kusuma School of Biological Science, Indian Institute of Technology (IIT), Delhi, India. 2.2. Synthesis 2.2.1. General method of synthesis of 2-amino-N-benzamides (3a-h; Scheme 1) Isatoic anhydride ( 1 ) (5–10 mmol) and solution of amine derivative 2(a-h) (5–10 mmol) were dissolved in 5–10 mL of dimenthylformamide (DMF) and refluxed at 50ºC for 8–10 hours. TLC (Ethanol: Chloroform) indicated the completion of reaction. TLC spots were visualized in UV Cabinet (HMG, India) (Fig. 3). The reaction mixture was brought to ambient temperature by adding cold deionized water. The precipitated solid was filtered and subsequently purified through recrystallization with ethanol to obtain the respective derivatives 3(a-h) [17]. 2.2.2. General method of synthesis of Quinazoline-2,4(1H,3H)-diones (4a-h; Scheme 2) Triphosgene (1 mmol) and potassium carbonate (5 mmol) were added to various substituted 2-amino-N-benzamides (0.84 mmol) in Tetrahydrofuran (THF) (20 ml) solution and refluxed for 15–20 hours. Later on, 50 mL of saturated NH 4 Cl solution was added to quench the reaction, and then (3 × 50 mL) portions of ethyl acetate were used for extraction. Mixed organic layers were rinsed with 40 mL of distilled water and anhydrous magnesium sulphate was used for drying. Crude solids (4a-h) were obtained by evaporating the solvent under vaccume [1]. The final compounds were recrystallized with ethanol. The reaction progress and purity of compounds was monitored through TLC by using ethyl acetate:chloroform (9:1) as the mobile phase. 2.2.2.1. 3-(4-Methylphenyl)quinazoline-2,4(1H,3H)-dione (4a) Yield = 79.58%; m.p.= 130-33 o C; IR(KBr): ν max = 3484 (NH Str, ArNH 2 ), 2925.7 (CH Str, CH 3 ), 2869.2 (C-H Str, Ar), 1719.5 (C = O, Str), 1690.5 (C = O Str, Cyclic CONH), 1455.7 (C = C Str, Ar), 1276.7 (C-N Str, Ar), 909.14 (NH, oop, wagging), 744.23 (C-H, oop, Ar) cm − 1 ; 1 H NMR (400MHz, DMSO- d 6 ) δ = 2.35 (s, 3H methyl ), 6.0 (s, 1H amide ), 7.0-7.6 (t, 4H, Ar-H), 7.75 (s, 1H benzene ), 8.13 (d, 1H benzene ), 8.2–8.4 (m, 2H benzene ) ppm; 13 C NMR (400MHz, DMSO-d 6 ) 100MHz δ = 24.3, 114.2, 115.8, 121.5, 123.3, 124.5, 127.7, 129.8, 132.4, 134.0, 137.9, 150.0, 158.9 ppm.; MS (m/z): 253.812 ([M + ]), 241.951, 230.000, 226.882, C 15 H 13 N 2 O 2 (253.81) 2.2.2.2. 3-(4-Methoxyphenyl)quinazoline-2,4(1H,3H)-dione (4b) Yield = 44.01%; m.p.= 162-165 o C; IR(KBr): ν max = 3481 (NH Str, ArNH 2 ), 2871 (C-H Str, Ar), 1720.5 (C = O, Str), 1690.5 (C = O Str, Cyclic CONH), 1455.7 (C = C Str, Ar), 1450.5 (CH, bending CH 3 ), 1275 (C-N Str, Ar), 909.14 (NH, oop, wagging), 742 (C-H, oop, Ar) cm − 1 , 1 H NMR (400MHz, DMSO- d 6 ) δ = 3.73 (s, 3H methoxy ), 6.02 (s, 1H amide ), 6.65 (s, 1H benzene ), 6.89 (s, 1H benzene ), 7.0-7.6 (t, 4H, Ar-H), 7.75 (s, 1H benzene ), 8.13 (d, 1H benzene ), 8.2–8.4 (m, 2H benzene ) ppm; 13 C NMR (400MHz, DMSO-d 6 ) 100MHz δ = 55.9, 114.5, 121.7, 123.3, 124.5, 125.1, 127.7, 132.4, 137.9, 150.0, 156.3, 158.9 ppm; MS (m/z): 269.953 ([M + ]), 242.055, 230.000 C 15 H 13 N 2 O 3 (269) 2.2.2.3. 3-(2-Aminophenyl)quinazoline-2,4(1H,3H)-dione (4c) Yield = 88.95%; m.p.= 233-234 o C; IR(KBr): ν max = 3479 (NH Str, ArNH 2 ), 2865 (C-H Str, Ar), 1724 (C = O, Str), 1690 (C = O Str, Cyclic CONH), 1650 (N-H, NH 2 bending) 1453 (C = C Str, Ar), 1352 (C-N Str, C-NH 2 ), 1282.5 (C-N Str, Ar), 909.14 (NH, oop, wagging), 741 (C-H, oop, Ar) cm − 1 ; 1 H NMR (400MHz, DMSO- d 6 ) δ = 4.01 (s, 2H amino ), 6.02 (s, 1H amide ), 6.43 (s, 1H benzene ), 6.76 (s, 1H benzene ), 7.0-7.6 (t, 4H, Ar-H), 7.75 (s, 1H benzene ), 8.13 (d, 1H benzene ), 8.2–8.4 (m, 2H benzene ) ppm; 13 CNMR (400 MHz, DMSO-d 6 ) 100MHz δ = 116.5, 119.0, 121.7, 123.3, 124.5, 126.8, 127.7, 132.4, 137.9, 141.8, 150.0, 158.9 ppm; MS (m/z): 253.036 ([M + ]), 251.025, 230.885, 223.506 C 14 H 11 N 3 O 2 (253) 2.2.2.4. 3-(3-Chloro-4-fluorophenyl)quinazoline-2,4(1H,3H)-dione (4d) Yield = 73.37%; m.p.= 160-163 o C; IR(KBr): ν max = 3490 (NH Str, ArNH 2 ), 2869.2 (C-H Str, Ar); 1724.3 (C = O, Str), 1690 (C = O Str, Cyclic CONH), 1448.3 (C = C Str, Ar), 1276.7 (C-N Str, Ar), 885.58 (N-H, oop, wagging), 740 (C-H, oop, Ar) cm − 1 ; 1 HNMR (400MHz, DMSO- d 6 ) δ = 6.02 (s, 1H amide ), 6.88 (d, 2H benzene ), 7.0-7.6 (t, 4H, Ar-H), 7.75 (s, 1H benzene ), 8.13 (d, 1H benzene ), 8.2–8.4 (m, 2H benzene ) ppm; 13 C NMR (400MHz, DMSO-d 6 ) 100MHz δ = 117.1, 121.0, 121.7, 123.3, 124.5, 127.7, 132.4, 137.9, 150.0, 158.4, 158.9 ppm; MS (m/z): 290.885 ([M + ]), 277.896, 252.885, 242.107, 230.862 C 14 H 8 N 2 O 2 ClF (290.5) 2.2.2.5. 3-(4-Nitrophenyl)quinazoline-2,4(1H,3H)-dione (4e) Yield = 69.03%; m.p.= 266-270 o C; IR(KBr): ν max = 3479 (NH Str, ArNH 2 ), 2864.5 (C-H Str, Ar), 1724.3 (C = O, Str), 1690 (C = O Str, Cyclic CONH), 1507.5, 1361.5 (N-O Str, NO 2 ), 1451 (C = C Str, Ar); 1281.4 (C-N Str, Ar), 899.72 (N-H, oop, wagging), 739 (C-H, oop, Ar) cm − 1 , 1 H NMR (400MHz, DMSO- d 6 ) δ = 6.02 (s, 1H amide ), 7.0-7.6 (t, 4H, Ar-H), 7.75 (s, 1H benzene ), 8.13 (d, 1H benzene ), 8.2–8.4 (m, 2H benzene ) ppm; 13 C NMR (400MHz, DMSO-d 6 ) 100MHz δ = 121.7, 122.5, 124.1, 127.7, 132.4, 137.9, 138.9, 143.6, 150.0, 158.9 ppm; MS (m/z): 283.090 ([M + ]), 278.235, 270.822, 261.022, 242.167, 238.805 C 14 H 9 N 3 O 4 (283) 2.2.2.6. 3-Cyclopropylquinazoline-2,4(1H,3H)-dione (4f) Yield = 90.85%; m.p.= 227-230 o C; IR(KBr): ν max = 3481 (NH Str, ArNH 2 ), 2925.7 (C-HStr, CH 2 ), 2864.5 (C-H Str, Ar), 1724.3 (C = O, Str), 1690 (C = O Str, Cyclic CONH), 1460.4 (C = C Str, Ar), 1352 (CH 2 deformation, cyclopropyl); 1083.5 (C-N Str, aliphatic); 880.87 (N-H, oop, wagging), 852.18 (CH 2 , cyclopropyl), 740 (C-H, oop, Ar) cm − 1 , 1 H NMR (400MHz, DMSO- d 6 ) δ = 0.31–0.56 (dd, 4H methylene ), 2.32 (s, 1H methane ), 6.0 (s, 1H amide ), 7.2–7.9 (t, 4H, ArH) ppm 13 C NMR (400MHz, DMSO-d 6 ) 100MHz δ = 4.4, 28.3, 121.7, 123.3, 124.5, 127.7, 132.4, 137.9, 151.5, 159.2 ppm; MS (m/z): 202.548 ([M + ]), 130.802 C 11 H 10 N 2 O 2 (202) 2.2.2.7. 3-(4-Bromophenyl)quinazoline-2,4(1H,3H)-dione (4g) Yield = 37.08%; m.p.= 240-243 o C; IR(KBr): ν max = 3479 (NH Str, ArNH 2 ), 2864 (C-H Str, Ar), 1725 (C = O, Str), 1690 (C = O Str, Cyclic CONH), 1452 (C = C Str, Ar), 1281 (C-N Str,Ar), 909.14 (NH, oop, wagging), 742 (C-H, oop, Ar), 680.5 (C-Br, Str) cm − 1 , 1 H NMR (400MHz, DMSO- d 6 ) δ = 6.0 (s, 1H amide ), 7.27 (t, 4H, ArH), 7.75 (s, 1H benzene ), 8.13 (d, 1H benzene ) ppm 13 C NMR (400MHz, DMSO-d 6 ) 100MHz δ = 118.7, 121.7, 123.8, 124.5, 127.7, 131.8, 132.4, 137.9, 150.0, 158.9 ppm; MS (m/z): 317.543 ([M + ]), 306.811, 291.002, 278.053, 260.811, 242.443, 233.506, 224.552 C 14 H 9 N 2 O 2 Br (316.9) 2.2.2.8. 3-(3-Nitrophenyl)quinazoline-2,4(1H,3H)-dione (4h) Yield = 79.49%; m.p.= 233-235 o C; IR(KBr): ν max = 3484 (NH Str, ArNH 2 ); 2897.5, 2869.2 (C-H Str, Ar); 1719.5 (C = O, Str); 1690 (C = O Str, Cyclic CONH); 1507.5, 1366.2 (N-O Str, NO 2 ); 1455 (C = C Str, Ar); 1281.4 (C-N Str, Ar); 885.58 (N-H, oop, wagging); 744.23 (C-H, oop, Ar) cm − 1 , 1 H NMR (400MHz, DMSO- d 6 ) δ = 6.0 (s, 1H amide ), 7.0-7.6 (t, 4H, ArH), 7.75 (s, 1H benzene ), 8.13 (d, 1H benzene ), 8.57 (s, 1H benzene ) ppm, 13 C NMR (400MHz, DMSO-d 6 ) 100MHz δ = 114.6, 119.5, 121.7, 123.3, 124.5, 127.7, 129.9, 132.4, 133.7, 137.9, 148.2, 150.0, 158.9 ppm; MS (m/z): 283.016 ([M + ]), 278.883, 265.441, 251.222, 244.698, 238.446, 225.832 C 14 H 9 N 3 O 4 (283) 2.3. Molecular Docking The inhibitory activity of the 3-Substituted quinazoline-2,4(1H,3H)-dione derivatives with respect to antibacterial, antifungal, and anticonvulsant activities was analyzed using a molecular docking strategy. The protein structures of interest, identified by their Protein Data Bank (PDB) accession codes 7VPR, 6BPP, and 2Z5X [18–20] were obtained from the RCSB Protein Data Bank repository in PDB file format. The active sites of each protein structure were pre-determined by the construction of a grid box around the co-crystallized ligand. The protein molecules were prepared using AutoDock Tools [21, 22]. Restoring residues, eliminating water, introducing polar hydrogens, and assigning Kollman charges were all steps in this procedure. The protein structures were subsequently converted to pdbqt format for further analysis. Three-dimensional conformations of the ligand molecules were generated using the Merck Molecular Force Field 94 (MMFF94) implemented in Auto Dock Vina. These 3D ligand structures were then exported and converted to pdbqt format utilizing Open Babel GUI. A Perl script was executed to facilitate the molecular docking of all ligand molecules against the aforementioned protein structures. The binding affinities, expressed as docking scores, were calculated for each ligand-receptor interaction and reported in kcal/mol. Pymol and Discovery Studio Visualizer was further used to gain insights into the molecular interaction study of the ligands with the best binding affinity against each protein. 2.4. Biological Evaluation 2.4. 1. Animals The anticonvulsant efficacy of compounds 4(a-h) was assessed using the maximal electroshock seizure (MES) model, alongside an evaluation of their neurotoxicity profile. The study employed Wistar strain albino rats (100–250 g) randomly allocated into three groups (n = 6 per group): control, standard, and test. Prior to and throughout the experimental period, subjects were maintained on a standard laboratory pellet diet (Hindustan Lever Ltd., India) with ad libitum access to water. Following group assignment, animals were allowed a 2–3 day acclimatization period in the novel environment before commencement of the experimental protocol. All procedures adhered strictly to the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) guidelines for the anticonvulsant screening of synthesized compounds. The study was reviewed and approval for the conducting of activity was given by the Institutional Animal Ethics Committee (IAEC Registration No-838/PO/Re/S/04/CPCSEA), Dr. K.N. Modi Institute of Pharmaceutical Education and Research, Modinagar, Uttar Pradesh, India. 2.4.2. Anticonvulsant Activity 2.4.2.1. Maximal electroshock seizure (MES) method All the synthesized compounds were subjected to MES method for anticonvulsant activity. Seizures were induced in all three groups by delivering electroshock of 60 Hz of alternating current 150mA for 0.2 sec by means of ear clip electrodes using electro-convulsiometer. 1% carboxymethylcellulose sodium (CMC) solution was used as control; phenytoin sodium (30mg/kg) was used as standard drug and administered through intraperitoneal route prior to evaluation. The dosage (30, 60, and 100 mg/kg body weight) were prepared as suspensions of synthesized compounds in 1% carboxymethylcellulose (CMC) and administered through intraperitoneal (i.p.) injection. Assessments were conducted at 0.5 and 4 hours post-administration. The anticonvulsant activity was quantified by observing the absence or reduction of the hind limb tonic extensor phase, a characteristic indicator of seizure severity in this model [13, 16, 23]. 2.4.2.2. Neurotoxicity Screening (Rotarod Method) Motor function assessment was conducted using the rotarod test to evaluate minimal motor impairment in the subjects. The apparatus consisted of a rotating rod with a diameter of 3.2 cm, initially set to a speed of 6 rpm with acceleration capability. Subjects were habituated to the device prior to testing. Following the training period, test compounds were administered intraperitoneally at a dose of 100 mg/kg body weight. Neurotoxicity was defined as the lack of ability to uphold balance on the rotating rod for atleast 60 seconds in each of four consecutive experiments. This method provided a quantitative measure of motor coordination and balance under the influence of the administered compounds [13, 16, 23]. 2.4.3 Antibacterial and Antifungal Activity The antimicrobial efficacy against bacterial and fungal pathogens was evaluated through in vitro assays using pure cultures. In vitro susceptibility testing was conducted employing the following methodologies: 2.4.3.1. Tube Dilution Method Dilutions of the synthesized compounds were prepared in growth medium to encompass their clinically relevant concentration ranges. Each tube, including a control without antimicrobial agent, was inoculated with an equivalent volume of broth containing 105–106 CFU/mL of bacteria. Following overnight incubation, the tubes were assessed for visible turbidity. Tube dilution method was resulted in the determination of antibacterial susceptibility in liquid media [24], and MIC determination for select derivatives. 2.4.3.2. Agar Diffusion Method Petri dishes containing agar medium were prepared using the pour plate method. The medium was subsequently inoculated with the target microorganisms. For the agar dilution assay, varying concentrations of antibiotics were incorporated into the agar medium to assess both aerobic and anaerobic microbial growth and incubated at 37°C for 24 hours. The antimicrobial agents diffused through the agar, resulting in clear zones of inhibition. The diameters of these zones were measured to estimate the efficacy of the antimicrobial substances [24]. Nutrient agar media was used for the purpose of preparation of agar plates for the growth of selected bacteria namely Gram positive strains- B. subtilis and S. aureus and Gram negative strains- E. coli and P. aeruginosa . Whereas Sabouraud’s agar media was used for the preparation of petri plates meant for the growth of fungi namely A. niger and C. albicans [25]. The Agar well diffusion test was performed by using nutrient agar medium as per the procedure given by Venkatesarlu et al. [26]. Each of the agar media-nutrient and Sabrouraud's was autoclaved for 15 minutes at 121ºC to ensure sterilityand then they were removed from the water bath immediately to cool them at 45ºC-55ºC [26]. Sterilization of petri dishes and pipettes was conducted in an oven at 150°C for 60 minutes. Subsequently, 25 mL of molten agar medium was aseptically dispensed into each sterilized Petri dish (10 cm diameter). To prepare inocula, bacterial cultures were propagated in 80 mL of nutrient broth, while fungal cultures were grown on Sabouraud's agar media. Both were incubated in 250 mL Erlenmeyer flasks. The inoculum flasks were incubated at 37ºC in case of bacteria for 18 hours and at 25ºC in case of fungi for 25 hours. Approximately 0.5 mL of inoculum broth containing various bacterial and fungal strains was introduced into separate petri dishes. A rotational technique was employed to ensure uniform distribution of the contents. Subsequently, the inoculated medium was allowed to solidify at ambient temperature. Standard drug used for antibacterial evaluation was ciprofloxacin and its stock solution was prepared using dissolving 1mg of drug in 1ml of dimethylsulfoxide (DMSO) to obtain 1000µg/mL of solution which was then serially diluted to obtain solutions of concentration 25, 50, 75, 100µg/mL. Griseofulvin was used as standard for the antifungal activity and its stock solution was prepared by dissolving 1mg of drug in 1mL of DMSO to obtain 1000µg/mL of solution which was then serially diluted to obtain solutions of concentration 25, 50, 75, 100µg/mL. All the test compounds were prepared by dissolving 1mg of drug in 1mL of DMSO to obtain 1000 µg/mL of solution which was then serially diluted to obtain solutions of concentration 25, 50, 75, 100 µg/mL [27]. 0.1 mL of the fraction of standard drug and test compounds were added in the wells and labeled accordingly. Petri-dishes were then kept in cool place for one hour to allow diffusion of solution into the medium and further incubated at 37ºC for 18 hours in case of bacteria and at 25ºC for 25 hours in case of fungi. Inhibition zones were observed surrounding the wells, and their diameters were measured in millimeters [26]. 3. Results and Discussion The compounds 3(a-h) and 4(a-h) were synthesized as per reactions given in the scheme 1 and scheme 2 respectively. 3.1. Molecular Docking To figure out the molecular mechanism behind the antibacterial activity of the 3-Substituted quinazoline-2,4 (1H,3H)-dione derivatives, a molecular docking approach was conducted against the Lipid A export ATP-binding/permease protein MsbA of gram-negative bacteria (PDB ID: 6BPP). Among the eight derivatives, compounds 4d and 4h showed the highest binding affinity i.e. -8.7 kcal/mol against MsbA protein. Compound 4d showed significant molecular interactions by demonstration of alkyl and pi-alkyl interaction at LEU171, LEU263, MET291, VAL178, ALA262, LEU294, pi-sigma interactions at LEU298 and ILE182, along with a halogen bond formation at ALA259. On the other hand compound 4h demonstrated pi-alkyl interactions at VAL178, ALA262, ALA259, and LEU171, along with pi-sigma bond formation at LEU298 and ILE182 (Fig. 4 , Table 1 ). To assess the antifungal activity of the synthesized compounds, a molecular docking was conducted against the fungal sterol uptake control protein 2 (Upc2) (PDB ID: 7VPR). Compound 4a exhibited highest binding affinity (-10 kcal/mol) against Upc2 protein. The significant molecular interactions of compound 4a were observed by demonstration of hydrogen bond formation at MET858, alkyl and pi-alkyl interactions at PRO836, VAL750, MET901, ARG859, and LEU906, pi-pi stacked bond formation at PHE905, and pi-sigma bond formation at VAL890 (Fig. 5 , Table 1 ). In-silico screening was also conducted against Human Monoamine Oxidase A (MAO-A) (PDB ID: 2Z5X) to gain insights of mechanism behind the anticonvulsant activity of aforesaid derivatives. Among all, compound 4h exhibited highest binding affinity of -10.7 kcal/mol against MAO-A protein. The significant interactions of compound 4h were demonstrated by hydrogen bond formation at TYR444, TYR69, ALA68, MET445, pi-donor hydrogen bond formation at GLY67 and TYR407, pi-pi stacked and pi-pi t-shaped bond formation at TYR444 and TYR407, along with pi-alkyl bond formation at ILE335 (Fig. 6 , Table 1 ). Table 1 Binding affinity scores of all the 3-Substituted quinazoline-2,4 (1H,3H)-dione derivatives against three different protein structures Compound Name Binding affinity against PDB ID:6BPP (kcal/mol) Binding affinity against PDB ID:7VPR (kcal/mol) Binding affinity against PDB ID:2Z5X (kcal/mol) 4a -8.4 -10 -9.6 4b -8.1 -9.6 -9.5 4c -8.2 -9.6 -9.3 4d -8.7 -9.9 -10.1 4e -8.5 -9.9 -10.2 4f -6.8 -8.4 -8.1 4g -8.4 -9.8 -9.6 4h -8.7 -9.6 -10.7 3.2. Anticonvulsant Activity The synthesized compounds 4 (a-h) demonstrated anticonvulsant activities which were evident through reduction in the duration of the hind limb tonic extensor phase (Table 2 ). Among all the compounds synthesized, compound 4h was found to be equipotent to the drug phenytoin sodium as it abolished seizures at the dose 30 mg/Kg at 0.5h and after 4h of administration. Compound 4d demonstrated anticonvulsant activity at a dose of 60 mg/kg, with seizure inhibition observed at both 0.5 and 4 hours post-administration.Compound 4a showed inhibited seizures at 60mg/kg at 0.5h and at 100mg/kg, after 4h of administration. Whereas compounds 4b , 4c , 4e , 4f and 4g inhibited seizures at 100mg/kg at 0.5h and after 4.0h of administration. Neurotoxicity screening was conducted on compounds 4(a-h) in which none of the synthesized compounds exhibited neurotoxic effects at a dose of 100 mg/kg when evaluated at 0.5 and 4 hours post-administration (Table 3 ). Table 2 Anticonvulsant activity of the synthesized compounds in the maximal electroshock seizure method Compound MES a 0.5h 4.0h 4a 4b 4c 4d 4e 4f 4g 4h Phenytoin b Sodium Control 1% (w/v)CMC c 60 100 100 60 100 100 100 30 30 1 100 100 100 60 100 100 100 30 30 1 a Intraperitoneal (i.p.) injections of the compound were administered to rats at doses of 30, 60, and 100 mg/kg. The values presented in the table represent the minimum dose at which bioactivity was observed in 50% or more of the test subjects. Examinations were conducted at 0.5 and 4.0 hours post-administration. b Reference drug phenytoin sodium (30mg/Kg). c Control compound 1% (w/v) CMC solution. Table 3 Evaluation of neurotoxic potential in the synthesized novel compounds Compound Neurotoxicity Screening a 0.5h 4.0h 4a 0/6 0/6 4b 0/6 0/6 4c 0/6 0/6 4d 0/6 0/6 4e 0/6 0/6 4f 0/6 0/6 4g 0/6 0/6 4h 0/6 0/6 Phenytoin b Sodium 0/6 0/6 a Dose of 100 mg/kg were administered through i.p injection in rat. Values indicate number of animals that showed neurotoxicity out of six animals 3.3. Antibacterial activity The in vitro antibacterial efficacy of novel compounds 4(a-h) was evaluated using the agar diffusion method. The assay was conducted against both Gram-positive (B. subtilis and S. aureus) and Gram-negative (E. coli and P. aeruginosa) bacterial strains. All compounds 4(a-h) were assessed for their antibacterial activity at a concentration of 100 µg/mL.The zone of inhibition (mm) was determined for each compound as measure of their activity along with Ciprofloxacin as standard drugs (Table 4 ). Outperforming all the other synthesised compounds, 4f showed strongest efficacy against all the selected bacteria, preventing their development at a concentration of 100 µg/mL (Fig. 7 ). However, all the synthesized compounds inhibited the growth of selected bacteria. Table 4 In vitro antibacterial activity of compounds 4(a-h) against selected strains Compound Zone of Inhibition S. aureus B . subtilis E . coli P. aeruginosa 4a 5.8 (50) 6.8 (50) 7.1 (50) 8.0 (50) 4b 9 (50) 11 (25) 5 (50) 7 (50) 4c 6 (25) 8 (12.5) 7 (50) 8 (25) 4d 5.2 (25) 5.9 (25) 8.3 (50) 8.7 (50) 4e 8.5 (25) 8.6 (25) 9.1(12.5) 9.4(12.5) 4f 12.1(6.25) 13.5(6.25) 11.8(12.5) 11.5 (12.5) 4g 6.6 (50) 6.8(25) 6.2(25) 7(6.25) 4h 8.1(25) 8.9(50) 7.5(100) 7.9(100) Ciprofloxacin 18(12.5) 19(6.25) 19(12.5) 17(6.25) Values in bracket are MIC values (µg/mL) 3.4. Antifungal activity The antifungal susceptibility of the developed compounds was evaluated using the agar diffusion method against the target bacteria viz . Fungus strains C. albicans and A. niger . The antifungal susceptibility of all synthesized compounds was evaluated at a concentration of 100 µg/mL using the agar diffusion method.The zone of inhibition (in mm) was determined for each compound as measure of their activity along with griseofulvin as standard drugs (Table 5 ). Among all the compounds synthesized, compound 4b showed maximum activity against selected fungal strains and it showed activity greater than the standard drug (Fig. 8 ). Compound 4d also showed greater activity but was less active than the standard drug. Table 5 In vitro antifungal activities of compounds 4(a-h) against selected strains Compound Zone of Inhibition C.albicans A.niger 4a 12 (100) 13 (100) 4b 25 (6.25) 25 (6.25) 4c 12 (100) 13 (100) 4d 21(25) 22 (6.25) 4e 19 (50) 18 (50) 4f 18 (25) 17 (50) 4g 19 (50) 19 (50) 4h 16 (100) 18 (100) Griseofulvin 23 (6.25) 23 (6.25) Values in bracket are MIC values (µg/mL) 4. Conclusion The main objective of this research was to synthesized a series of quinazoline-2,4-dione derivatives with medicinal activity and minimal adverse effects.The therapeutic potential of 3-substituted quinazoline-2,4(1H,3H)-diones derivatives may be the new vista of research there in this particular area.A series of 3-substituted quinazoline2,4(1H,3H)-dione derivatives was prepared starting from reaction of isatoic anhydride with different aromatic amines by taking dimethylformamide as solvent to yield 2-amino-N-benzamide derivatives in first scheme followed by the reaction with triphosgene in the presence of potassium carbonate by taking tetrahydrofuran to bring about cyclization to finally produce 3-substituted-quinazoline-2,4(1H,3H)-dione derivatives in second scheme. All the synthesized compounds were purified by re-crystallization and identified on the basis of physicochemical characterization melting point and spectral characterization by FTIR, H 1 NMR, 13 CNMR and MALDI-MS spectral method. In vitro evaluation using the MES method revealed significant anticonvulsant activity of the synthesized compounds compared to the standard drug, phenytoin sodium. However, all of the synthesized compounds also exhibited antibacterial and anti-fungal activities when compared with the standard drugs- Ciprofloxacin and Griseofulvin for antimicrobial screening purpose. Compound 4h was found to be equipotent to the drug phenytoin sodium which makes us conclude that it may serve as lead for the synthesis of new compounds with anticonvulsant action. Compound 4f was effective against all the selected bacteria and inhibited their growth so it can be used further for the synthesis of compounds with great antibacterial activity. Compound 4b exhibited activity greater than the standard drug griseofulvin therefore; it may serve as lead for the synthesis of series of new compounds with greater antifungal activity. Abbreviations 1 H NMR; Proton Nuclear Magnetic Resonance, AMPA; α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid, A. niger; Aspergillus niger, ATP; Adenosine Triphosphate, B. Subtilis; Bacillus subtilis, C. albicans; Candida albicans, DMEM; Dulbecco's Modified Eagle's Media, DMF; Dimethylformamide, DMSO; Dimethyl Sulfoxide, DNA; Deoxyribonucleic Acid, E. coli ; Escherichia coli, ERG; Electron Releasing Group, EWG; Electron-Withdrawing Group, FT-IR; Fourier-Transform Infrared Spectroscopy, IC 50 ; Half-Maximal Inhibitory Concentration, HCl; Hydrochloric Acid, m; Meta, o; Ortho, p; Para, PBS; Phosphate-Buffered Saline, PCT; Proximal Convoluted Tubule, P. aeruginosa; Pseudomonas aeruginosa, ppm; parts per million, RBF; Round Bottom Flask, SAR; Structure-activity Relationship, S. aureus ; Staphylococcus aureus , THF; Tetrahydrofuran, TLC; Thin-Layer Chromatography, TPSA; Total Polar Surface Area Declarations Acknowledgement The synthetic work and biological activity supported by Dr. KNMIPER, Modinagar, Uttar Pradesh, India is greatly acknowledged. Authors duly acknowledge Kusuma School of Biological Science, Indian Institute of Technology (IIT), Delhi, India for providing MALDI-MS analysis. Authors are also thankful to Sophisticated Analytical Instrument Facility (SAIF), Panjab University, Chandigarh, India for providing spectroscopic facilities. Data availability The authors declare that the data supporting the findings of this study are available within the paper and its Supplementary Information files. Should any raw data files be needed in another format they are available from the corresponding author upon reasonable request. Source data are provided with this paper. Author Contributions Conceptualization and study design were carried out by the corresponding author. Chemical synthesis and characterization of the compounds were performed by Neelam Sharma. Biological evaluation, including anticonvulsant, antibacterial, and antifungal studies, was conducted by Manish Kumar and Vijay Kumar Sharma. Molecular docking studies and computational analysis were performed by Ratul Bhowmik and Corresponding author. Data analysis and interpretation were carried out by Manish Kumar. The manuscript was written by Neelam Sharma and corresponding author and reviewed and approved by all authors. Conflict of Interests The authors report no conflict of interests. References Li X, Lee YR. Facile One-Pot Synthesis of Quinazoline-2,4-dione Derivatives and Application to Naturally Occurring Alkaloids from Zanthoxylum Arborescens. Bulletin of the Korean Chemical Society. 2011;32:2121–2124 Jin HZ, Du JL, Zhang WD, Yan SK, Chen HS, Lee JH, Lee JJ. A new quinazolinedione alkaloid from the fruits of Evodia officinalis. Fitoterapia. 2008;79:317–318 Michael, JP. Quinoline, quinazoline and acridone alkaloids. Nat. Prod. Rep. 2001;18: 543–559 ChawlaA,BatraC. Recent advances of quinazolinone derivatives as marker for various biological activities. Int. Res. J. Pharm. 2013;4:49–58 T Panneerselvam, Kumar PV. 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Synthesis and Biological Evaluation of 2-Aminobenzamide Derivatives as Antimicrobial Agents: Opening/Closing Pharmacophore Site. International Journal of Molecular Sciences. 2014;15:5115–5127 Elalouf A. In-silico Structural Modeling of Human Immunodeficiency Virus Proteins. Biomed Eng Comput Biol. 2023;14:11795972231154402 Ho H, Miu A, Alexander MK, Garcia NK, Oh A, Zilberleyb I, Reichelt M, Austin CD, Tam C, Shriver S, Hu H, Labadie SS, Liang J, Wang L, Wang J, Lu Y, PurkeyHE, Quinn J, Franke Y, Clark K, Beresini MH, Tan MW, Sellers BD, Maurer T, Koehler MFT, Wecksler AT, Kiefer JR, Verma V, Xu Y, Nishiyama M, Payandeh J, Koth CM. Structural basis for dual-mode inhibition of the ABC transporter MsbA. Nature. 2018;557:196–201 Son SY, Ma J, Kondou Y, Yoshimura M, Yamashita E, Tsukihara T. Structure of human monoamine oxidase A at 2.2-A resolution: the control of opening the entry for substrates/inhibitors. Proc Natl Acad Sci U S A. 2008;105:5739–44 EberhardtJ, Santos-Martins D, Tillack AF, Forli S. AutoDock Vina 1.2.0: New Docking Methods, Expanded Force Field, and Python Bindings. J. Chem. Inf. Model. 2021;61:3891–3898 Tanchuk VY, Tanin VO, Vovk AI, Poda, G. A New, improved hybrid scoring function for Molecular Docking and scoring based on AutoDock and AutoDock Vina. Chem Biol Drug Des. 2016;87:618–625 KulkarniSK. Handbook of Experimental Pharmacology (3rd ed), Vallabh Prakashan, Delhi,1999;Reprint 2009:131–134 Indian Pharmacopoeia commission, Govt. of India. Micrbiological Assay of Antibiotics, Indian Pharmacopoeia, 2014;3(7):38–50 Black WD. A comparison of several media types and basic techniques used to assess outdoor airborne fungi in Melbourne, Australia. PLoS One. 2020;15:e0238901 Venkatesarulu V, Kokate C,Rambhau D, Ciddi V. Antimicrobial activity of chemoconstituents of roots of salaciamacrosperma. Ancient science of life. 1992;12: 251–256 Zaranappa, Vagdevi HM, Jayanna ND, Latha KP. Synthesis, Characterization and evaluation of Antibacterial Activity of some 3-Substitutedphenylquinazoline – 2,4-diones. Der Pharma Chem. 2012;4: 1754–1758 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-8566641\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":true,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":593468827,\"identity\":\"32d2b69d-7e34-4ac4-8ffe-d591514d73c5\",\"order_by\":0,\"name\":\"Neelam Sharma\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Shri Ram College of Pharmacy\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Neelam\",\"middleName\":\"\",\"lastName\":\"Sharma\",\"suffix\":\"\"},{\"id\":593468828,\"identity\":\"0a62e3dc-5523-4e12-a488-180669dddaf7\",\"order_by\":1,\"name\":\"Nishtha Shalmali\",\"email\":\"data:image/png;base64,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\",\"orcid\":\"\",\"institution\":\"Dr. K. 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Ar = Aryl hydrophobic binding site, HBD = A hydrogen bonding domain, D = An electron donor system\\u003c/strong\\u003e\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8566641/v1/9c4880eea91c29c094e47c2d.png\"},{\"id\":103505204,\"identity\":\"6dceb7f0-4322-493c-a3c8-b6b66a901e5c\",\"added_by\":\"auto\",\"created_at\":\"2026-02-26 13:27:43\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":71479,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eScheme for the synthesis of 2-amino-N-benzamide derivatives from Isatoic Anhydride and amine derivatives in DMF (Scheme1) and scheme for the synthesis of 3-Substitutedquinazoline-2,4(1H,3H)-dione derivatives from 2-amino-N-benzamide derivatives by cyclization with triphogene (Scheme 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6BPP\\u003c/strong\\u003e\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"4.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8566641/v1/ef70b745a50e9622a74885c3.png\"},{\"id\":103174547,\"identity\":\"423f10c7-7b54-4e17-bbad-4d42a242a911\",\"added_by\":\"auto\",\"created_at\":\"2026-02-22 15:49:49\",\"extension\":\"png\",\"order_by\":5,\"title\":\"Figure 5\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":243657,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003e(i) 3D representation of molecular interaction between compound 4a and protein 7VPR (ii) 2D representation of molecular interaction between compound 4a and protein 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2Z5X\\u003c/strong\\u003e\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"6.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8566641/v1/c9e4de5f6996ea26c743c70a.png\"},{\"id\":103174544,\"identity\":\"b38571ca-1cad-4c4e-8f51-320e7413efb4\",\"added_by\":\"auto\",\"created_at\":\"2026-02-22 15:49:49\",\"extension\":\"png\",\"order_by\":7,\"title\":\"Figure 7\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":204472,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eZones observed during in vitro antibacterial activity of compound 4f\\u003c/strong\\u003e\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"7.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8566641/v1/82874ac4d0dd5c9bb19b1e67.png\"},{\"id\":103504384,\"identity\":\"25e29f79-0b10-4c67-8142-fd5c8cd69a48\",\"added_by\":\"auto\",\"created_at\":\"2026-02-26 13:19:38\",\"extension\":\"png\",\"order_by\":8,\"title\":\"Figure 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Bioactivity: Synthesis and Pharmacological Profiling of 3-Substituted Quinazoline-2,4-dione Heterocycles\",\"fulltext\":[{\"header\":\"1. Introduction\",\"content\":\"\\u003cp\\u003e \\u003cdiv class=\\\"BlockQuote\\\"\\u003e \\u003cp\\u003eQuinazolinedione is a heterocyclic moiety present in many naturally occurring alkaloids isolated from plant species like \\u003cem\\u003eZanthoxylum arborescens\\u003c/em\\u003e [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e], \\u003cem\\u003eEvodia officinalis\\u003c/em\\u003e [\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e] and from Phylloperthadiversa (chafer beetle) [\\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e]. Many synthetic drugs like ketanserin (anti-hypertensive) [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e], benzouracil (anti-viral) [\\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e], selurampanel (anti-convulsant) [\\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e6\\u003c/span\\u003e], etc. are stated to contain quinazolinedione moiety (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e).\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"BlockQuote\\\"\\u003e \\u003cp\\u003eMoreover, quinazolinedione derivatives reported for activities like anti-inflammatory [\\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e], anti-serotonergic [\\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e], anti-microbial [\\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e], anti-cancer [\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e10\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e] etc. It is evident that quinazolinediones act as anticonvulsant by competitively binding to glutamate receptor, kainate receptor and AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid) receptor. This leads to reduction in glutamate binding affinity for the receptors and sodium ion channels remain closed and thus inhibition of seizures [\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e]. The literature survey revealed that quinazolinedione moiety if present in any compound shows anticonvulsant action due to following features mentioned in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e [\\u003cspan additionalcitationids=\\\"CR14 CR15\\\" citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e16\\u003c/span\\u003e]: hydrophobic aryl domain (Ar), hydrogen bonding domain (HBD) in the form of \\u0026ndash;NHCO- grouping and electron donor atom (D).\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eBased on these features, a series of 3-substitutedquinazoline-2,4(1H,3H)-dione analogues were synthesized and screened for anti-convulsant activity against maximal electroshock (MES) convulsions. These derivatives were also screened for anti-bacterial and anti-fungal activity against selected bacterial and fungal strains.\\u003c/p\\u003e\"},{\"header\":\"2. Material and Methods\",\"content\":\"\\u003cdiv id=\\\"Sec3\\\"\\u003e\\n \\u003ch2\\u003e2.1. Chemistry\\u003c/h2\\u003e\\n \\u003cp\\u003eIsatoic anhydride and triphosgene were acquired from Sigma-Aldrich (India), while remaining reagents were sourced from Merck which is located in India and Central Drug House (Delhi, India). A digital melting point device was utilised in order to ascertain the melting points of each and every compound that was synthesized. Silica Gel GF 256 (Merck) was used to coat TLC plates and TLC spots of synthesized compounds were visualized using TLC UV Cabinet (HMG, India). The \\u003csup\\u003e1\\u003c/sup\\u003eHNMR and \\u003csup\\u003e13\\u003c/sup\\u003eCNMR spectra were recorded on a Bruker Advance II NMR (400 MHz) Spectrometer in DMSO-d\\u003csub\\u003e6\\u003c/sub\\u003e at Punjab University, Chandigarh, Punjab, India. IR spectra were recorded on a Perkin Elmer FTIR 1650 Spectrometer. Mass spectra were recorded on Bruker MALDI Ultra Flextreme using 2,5-dihydroxybenzoic acid (DHB) matrix at Kusuma School of Biological Science, Indian Institute of Technology (IIT), Delhi, India.\\u003c/p\\u003e\\n\\u003c/div\\u003e\\n\\u003cdiv id=\\\"Sec4\\\"\\u003e\\n \\u003ch2\\u003e2.2. Synthesis\\u003c/h2\\u003e\\n \\u003cdiv id=\\\"Sec5\\\"\\u003e\\n \\u003ch2\\u003e2.2.1. General method of synthesis of 2-amino-N-benzamides (3a-h; Scheme 1)\\u003c/h2\\u003e\\n \\u003cp\\u003eIsatoic anhydride (\\u003cstrong\\u003e1\\u003c/strong\\u003e) (5–10 mmol) and solution of amine derivative \\u003cstrong\\u003e2(a-h)\\u003c/strong\\u003e (5–10 mmol) were dissolved in 5–10 mL of dimenthylformamide (DMF) and refluxed at 50ºC for 8–10 hours. TLC (Ethanol: Chloroform) indicated the completion of reaction. TLC spots were visualized in UV Cabinet (HMG, India) (Fig. 3). The reaction mixture was brought to ambient temperature by adding cold deionized water. The precipitated solid was filtered and subsequently purified through recrystallization with ethanol to obtain the respective derivatives \\u003cstrong\\u003e3(a-h)\\u003c/strong\\u003e[17].\\u003c/p\\u003e\\n \\u003c/div\\u003e\\n \\u003cdiv id=\\\"Sec6\\\"\\u003e\\n \\u003ch2\\u003e2.2.2. General method of synthesis of Quinazoline-2,4(1H,3H)-diones (4a-h; Scheme 2)\\u003c/h2\\u003e\\n \\u003cp\\u003eTriphosgene (1 mmol) and potassium carbonate (5 mmol) were added to various substituted 2-amino-N-benzamides (0.84 mmol) in Tetrahydrofuran (THF) (20 ml) solution and refluxed for 15–20 hours. Later on, 50 mL of saturated NH\\u003csub\\u003e4\\u003c/sub\\u003eCl solution was added to quench the reaction, and then (3 × 50 mL) portions of ethyl acetate were used for extraction. Mixed organic layers were rinsed with 40 mL of distilled water and anhydrous magnesium sulphate was used for drying. Crude solids (4a-h) were obtained by evaporating the solvent under vaccume [1]. The final compounds were recrystallized with ethanol. The reaction progress and purity of compounds was monitored through TLC by using ethyl acetate:chloroform (9:1) as the mobile phase.\\u003c/p\\u003e\\n \\u003cdiv id=\\\"Sec7\\\"\\u003e\\n \\u003ch2\\u003e\\u003cstrong\\u003e2.2.2.1.\\u003c/strong\\u003e 3-(4-Methylphenyl)quinazoline-2,4(1H,3H)-dione \\u003cstrong\\u003e(4a)\\u003c/strong\\u003e\\u003c/h2\\u003e\\n \\u003cp\\u003eYield = 79.58%; m.p.= 130-33\\u003csup\\u003eo\\u003c/sup\\u003eC; IR(KBr): ν\\u003csub\\u003emax\\u003c/sub\\u003e = 3484 (NH Str, ArNH\\u003csub\\u003e2\\u003c/sub\\u003e), 2925.7 (CH Str, CH\\u003csub\\u003e3\\u003c/sub\\u003e), 2869.2 (C-H Str, Ar), 1719.5 (C = O, Str), 1690.5 (C = O Str, Cyclic CONH), 1455.7 (C = C Str, Ar), 1276.7 (C-N Str, Ar), 909.14 (NH, oop, wagging), 744.23 (C-H, oop, Ar) cm\\u003csup\\u003e− 1\\u003c/sup\\u003e; \\u003csup\\u003e1\\u003c/sup\\u003eH NMR (400MHz, DMSO-\\u003cem\\u003ed\\u003c/em\\u003e\\u003csub\\u003e\\u003cem\\u003e6\\u003c/em\\u003e\\u003c/sub\\u003e) \\u003cem\\u003eδ\\u003c/em\\u003e = 2.35 (s, 3H \\u003csub\\u003emethyl\\u003c/sub\\u003e), 6.0 (s, 1H \\u003csub\\u003eamide\\u003c/sub\\u003e), 7.0-7.6 (t, 4H, Ar-H), 7.75 (s, 1H \\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.13 (d, 1H \\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.2–8.4 (m, 2H\\u003csub\\u003ebenzene\\u003c/sub\\u003e) ppm; \\u003csup\\u003e13\\u003c/sup\\u003eC NMR (400MHz, DMSO-d\\u003csub\\u003e6\\u003c/sub\\u003e) 100MHz \\u003cem\\u003eδ\\u003c/em\\u003e = 24.3, 114.2, 115.8, 121.5, 123.3, 124.5, 127.7, 129.8, 132.4, 134.0, 137.9, 150.0, 158.9 ppm.; MS (m/z): 253.812 ([M\\u003csup\\u003e+\\u003c/sup\\u003e]), 241.951, 230.000, 226.882, C\\u003csub\\u003e15\\u003c/sub\\u003eH\\u003csub\\u003e13\\u003c/sub\\u003e N\\u003csub\\u003e2\\u003c/sub\\u003eO\\u003csub\\u003e2\\u003c/sub\\u003e (253.81)\\u003c/p\\u003e\\n \\u003cdiv id=\\\"Sec8\\\"\\u003e\\n \\u003ch2\\u003e\\u003cstrong\\u003e2.2.2.2.\\u003c/strong\\u003e\\u003cem\\u003e3-(4-Methoxyphenyl)quinazoline-2,4(1H,3H)-dione\\u003c/em\\u003e \\u003cstrong\\u003e(4b)\\u003c/strong\\u003e\\u003c/h2\\u003e\\n \\u003cp\\u003eYield = 44.01%; m.p.= 162-165\\u003csup\\u003eo\\u003c/sup\\u003eC; IR(KBr): ν\\u003csub\\u003emax\\u003c/sub\\u003e = 3481 (NH Str, ArNH\\u003csub\\u003e2\\u003c/sub\\u003e), 2871 (C-H Str, Ar), 1720.5 (C = O, Str), 1690.5 (C = O Str, Cyclic CONH), 1455.7 (C = C Str, Ar), 1450.5 (CH, bending CH\\u003csub\\u003e3\\u003c/sub\\u003e), 1275 (C-N Str, Ar), 909.14 (NH, oop, wagging), 742 (C-H, oop, Ar) cm\\u003csup\\u003e− 1\\u003c/sup\\u003e, \\u003csup\\u003e1\\u003c/sup\\u003eH NMR (400MHz, DMSO-\\u003cem\\u003ed\\u003c/em\\u003e\\u003csub\\u003e\\u003cem\\u003e6\\u003c/em\\u003e\\u003c/sub\\u003e) \\u003cem\\u003eδ\\u003c/em\\u003e = 3.73 (s, 3H \\u003csub\\u003emethoxy\\u003c/sub\\u003e), 6.02 (s, 1H \\u003csub\\u003eamide\\u003c/sub\\u003e), 6.65 (s, 1H \\u003csub\\u003ebenzene\\u003c/sub\\u003e), 6.89 (s, 1H \\u003csub\\u003ebenzene\\u003c/sub\\u003e), 7.0-7.6 (t, 4H, Ar-H), 7.75 (s, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.13 (d, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.2–8.4 (m, 2H\\u003csub\\u003ebenzene\\u003c/sub\\u003e) ppm; \\u003csup\\u003e13\\u003c/sup\\u003eC NMR (400MHz, DMSO-d\\u003csub\\u003e6\\u003c/sub\\u003e) 100MHz \\u003cem\\u003eδ\\u003c/em\\u003e = 55.9, 114.5, 121.7, 123.3, 124.5, 125.1, 127.7, 132.4, 137.9, 150.0, 156.3, 158.9 ppm; MS (m/z): 269.953 ([M\\u003csup\\u003e+\\u003c/sup\\u003e]), 242.055, 230.000 C\\u003csub\\u003e15\\u003c/sub\\u003eH\\u003csub\\u003e13\\u003c/sub\\u003e N\\u003csub\\u003e2\\u003c/sub\\u003eO\\u003csub\\u003e3\\u003c/sub\\u003e (269)\\u003c/p\\u003e\\n \\u003c/div\\u003e\\n \\u003cdiv id=\\\"Sec9\\\"\\u003e\\n \\u003ch2\\u003e\\u003cstrong\\u003e2.2.2.3.\\u003c/strong\\u003e\\u003cem\\u003e3-(2-Aminophenyl)quinazoline-2,4(1H,3H)-dione\\u003c/em\\u003e \\u003cstrong\\u003e(4c)\\u003c/strong\\u003e\\u003c/h2\\u003e\\n \\u003cp\\u003eYield = 88.95%; m.p.= 233-234\\u003csup\\u003eo\\u003c/sup\\u003eC; IR(KBr): ν\\u003csub\\u003emax\\u003c/sub\\u003e = 3479 (NH Str, ArNH\\u003csub\\u003e2\\u003c/sub\\u003e), 2865 (C-H Str, Ar), 1724 (C = O, Str), 1690 (C = O Str, Cyclic CONH), 1650 (N-H, NH\\u003csub\\u003e2\\u003c/sub\\u003e bending) 1453 (C = C Str, Ar), 1352 (C-N Str, C-NH\\u003csub\\u003e2\\u003c/sub\\u003e), 1282.5 (C-N Str, Ar), 909.14 (NH, oop, wagging), 741 (C-H, oop, Ar) cm\\u003csup\\u003e− 1\\u003c/sup\\u003e ; \\u003csup\\u003e1\\u003c/sup\\u003eH NMR (400MHz, DMSO-\\u003cem\\u003ed\\u003c/em\\u003e\\u003csub\\u003e\\u003cem\\u003e6\\u003c/em\\u003e\\u003c/sub\\u003e) \\u003cem\\u003eδ\\u003c/em\\u003e = 4.01 (s, 2H\\u003csub\\u003eamino\\u003c/sub\\u003e), 6.02 (s, 1H\\u003csub\\u003eamide\\u003c/sub\\u003e), 6.43 (s, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 6.76 (s, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 7.0-7.6 (t, 4H, Ar-H), 7.75 (s, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.13 (d, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.2–8.4 (m, 2H\\u003csub\\u003ebenzene\\u003c/sub\\u003e) ppm; \\u003csup\\u003e13\\u003c/sup\\u003eCNMR (400 MHz, DMSO-d\\u003csub\\u003e6\\u003c/sub\\u003e) 100MHz \\u003cem\\u003eδ\\u003c/em\\u003e = 116.5, 119.0, 121.7, 123.3, 124.5, 126.8, 127.7, 132.4, 137.9, 141.8, 150.0, 158.9 ppm; MS (m/z): 253.036 ([M\\u003csup\\u003e+\\u003c/sup\\u003e]), 251.025, 230.885, 223.506 C\\u003csub\\u003e14\\u003c/sub\\u003eH\\u003csub\\u003e11\\u003c/sub\\u003eN\\u003csub\\u003e3\\u003c/sub\\u003eO\\u003csub\\u003e2\\u003c/sub\\u003e (253)\\u003c/p\\u003e\\n \\u003c/div\\u003e\\n \\u003cdiv id=\\\"Sec10\\\"\\u003e\\n \\u003ch2\\u003e\\u003cstrong\\u003e2.2.2.4.\\u003c/strong\\u003e\\u003cem\\u003e3-(3-Chloro-4-fluorophenyl)quinazoline-2,4(1H,3H)-dione\\u003c/em\\u003e \\u003cstrong\\u003e(4d)\\u003c/strong\\u003e\\u003c/h2\\u003e\\n \\u003cp\\u003eYield = 73.37%; m.p.= 160-163\\u003csup\\u003eo\\u003c/sup\\u003eC; IR(KBr): ν\\u003csub\\u003emax\\u003c/sub\\u003e = 3490 (NH Str, ArNH\\u003csub\\u003e2\\u003c/sub\\u003e), 2869.2 (C-H Str, Ar); 1724.3 (C = O, Str), 1690 (C = O Str, Cyclic CONH), 1448.3 (C = C Str, Ar), 1276.7 (C-N Str, Ar), 885.58 (N-H, oop, wagging), 740 (C-H, oop, Ar) cm\\u003csup\\u003e− 1\\u003c/sup\\u003e; \\u003csup\\u003e1\\u003c/sup\\u003eHNMR (400MHz, DMSO-\\u003cem\\u003ed\\u003c/em\\u003e\\u003csub\\u003e\\u003cem\\u003e6\\u003c/em\\u003e\\u003c/sub\\u003e) \\u003cem\\u003eδ\\u003c/em\\u003e = 6.02 (s, 1H\\u003csub\\u003eamide\\u003c/sub\\u003e), 6.88 (d, 2H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 7.0-7.6 (t, 4H, Ar-H), 7.75 (s, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.13 (d, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.2–8.4 (m, 2H\\u003csub\\u003ebenzene\\u003c/sub\\u003e) ppm; \\u003csup\\u003e13\\u003c/sup\\u003eC NMR (400MHz, DMSO-d\\u003csub\\u003e6\\u003c/sub\\u003e) 100MHz \\u003cem\\u003eδ\\u003c/em\\u003e = 117.1, 121.0, 121.7, 123.3, 124.5, 127.7, 132.4, 137.9, 150.0, 158.4, 158.9 ppm; MS (m/z): 290.885 ([M\\u003csup\\u003e+\\u003c/sup\\u003e]), 277.896, 252.885, 242.107, 230.862 C\\u003csub\\u003e14\\u003c/sub\\u003eH\\u003csub\\u003e8\\u003c/sub\\u003eN\\u003csub\\u003e2\\u003c/sub\\u003eO\\u003csub\\u003e2\\u003c/sub\\u003eClF (290.5)\\u003c/p\\u003e\\n \\u003c/div\\u003e\\n \\u003cdiv id=\\\"Sec11\\\"\\u003e\\n \\u003ch2\\u003e\\u003cstrong\\u003e2.2.2.5.\\u003c/strong\\u003e\\u003cem\\u003e3-(4-Nitrophenyl)quinazoline-2,4(1H,3H)-dione\\u003c/em\\u003e \\u003cstrong\\u003e(4e)\\u003c/strong\\u003e\\u003c/h2\\u003e\\n \\u003cp\\u003eYield = 69.03%; m.p.= 266-270\\u003csup\\u003eo\\u003c/sup\\u003eC; IR(KBr): ν\\u003csub\\u003emax\\u003c/sub\\u003e = 3479 (NH Str, ArNH\\u003csub\\u003e2\\u003c/sub\\u003e), 2864.5 (C-H Str, Ar), 1724.3 (C = O, Str), 1690 (C = O Str, Cyclic CONH), 1507.5, 1361.5 (N-O Str, NO\\u003csub\\u003e2\\u003c/sub\\u003e), 1451 (C = C Str, Ar); 1281.4 (C-N Str, Ar), 899.72 (N-H, oop, wagging), 739 (C-H, oop, Ar) cm\\u003csup\\u003e− 1\\u003c/sup\\u003e, \\u003csup\\u003e1\\u003c/sup\\u003eH NMR (400MHz, DMSO-\\u003cem\\u003ed\\u003c/em\\u003e\\u003csub\\u003e\\u003cem\\u003e6\\u003c/em\\u003e\\u003c/sub\\u003e) \\u003cem\\u003eδ\\u003c/em\\u003e = 6.02 (s, 1H\\u003csub\\u003eamide\\u003c/sub\\u003e), 7.0-7.6 (t, 4H, Ar-H), 7.75 (s, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.13 (d, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.2–8.4 (m, 2H\\u003csub\\u003ebenzene\\u003c/sub\\u003e) ppm; \\u003csup\\u003e13\\u003c/sup\\u003eC NMR (400MHz, DMSO-d\\u003csub\\u003e6\\u003c/sub\\u003e) 100MHz \\u003cem\\u003eδ\\u003c/em\\u003e = 121.7, 122.5, 124.1, 127.7, 132.4, 137.9, 138.9, 143.6, 150.0, 158.9 ppm; MS (m/z): 283.090 ([M\\u003csup\\u003e+\\u003c/sup\\u003e]), 278.235, 270.822, 261.022, 242.167, 238.805 C\\u003csub\\u003e14\\u003c/sub\\u003eH\\u003csub\\u003e9\\u003c/sub\\u003eN\\u003csub\\u003e3\\u003c/sub\\u003eO\\u003csub\\u003e4\\u003c/sub\\u003e (283)\\u003c/p\\u003e\\n \\u003c/div\\u003e\\n \\u003c/div\\u003e\\n \\u003cdiv id=\\\"Sec12\\\"\\u003e\\n \\u003ch2\\u003e\\u003cstrong\\u003e2.2.2.6.\\u003c/strong\\u003e \\u003cem\\u003e3-Cyclopropylquinazoline-2,4(1H,3H)-dione\\u003c/em\\u003e \\u003cstrong\\u003e(4f)\\u003c/strong\\u003e\\u003c/h2\\u003e\\n \\u003cp\\u003eYield = 90.85%; m.p.= 227-230\\u003csup\\u003eo\\u003c/sup\\u003eC; IR(KBr): ν\\u003csub\\u003emax\\u003c/sub\\u003e = 3481 (NH Str, ArNH\\u003csub\\u003e2\\u003c/sub\\u003e), 2925.7 (C-HStr, CH\\u003csub\\u003e2\\u003c/sub\\u003e), 2864.5 (C-H Str, Ar), 1724.3 (C = O, Str), 1690 (C = O Str, Cyclic CONH), 1460.4 (C = C Str, Ar), 1352 (CH\\u003csub\\u003e2\\u003c/sub\\u003e deformation, cyclopropyl); 1083.5 (C-N Str, aliphatic); 880.87 (N-H, oop, wagging), 852.18 (CH\\u003csub\\u003e2\\u003c/sub\\u003e, cyclopropyl), 740 (C-H, oop, Ar) cm\\u003csup\\u003e− 1\\u003c/sup\\u003e, \\u003csup\\u003e1\\u003c/sup\\u003eH NMR (400MHz, DMSO-\\u003cem\\u003ed\\u003c/em\\u003e\\u003csub\\u003e\\u003cem\\u003e6\\u003c/em\\u003e\\u003c/sub\\u003e) \\u003cem\\u003eδ\\u003c/em\\u003e = 0.31–0.56 (dd, 4H\\u003csub\\u003emethylene\\u003c/sub\\u003e), 2.32 (s, 1H\\u003csub\\u003emethane\\u003c/sub\\u003e), 6.0 (s, 1H\\u003csub\\u003eamide\\u003c/sub\\u003e), 7.2–7.9 (t, 4H, ArH) ppm \\u003csup\\u003e13\\u003c/sup\\u003eC NMR (400MHz, DMSO-d\\u003csub\\u003e6\\u003c/sub\\u003e) 100MHz \\u003cem\\u003eδ\\u003c/em\\u003e = 4.4, 28.3, 121.7, 123.3, 124.5, 127.7, 132.4, 137.9, 151.5, 159.2 ppm; MS (m/z): 202.548 ([M\\u003csup\\u003e+\\u003c/sup\\u003e]), 130.802 C\\u003csub\\u003e11\\u003c/sub\\u003eH\\u003csub\\u003e10\\u003c/sub\\u003e N\\u003csub\\u003e2\\u003c/sub\\u003eO\\u003csub\\u003e2\\u003c/sub\\u003e (202)\\u003c/p\\u003e\\n \\u003cdiv id=\\\"Sec13\\\"\\u003e\\n \\u003ch2\\u003e\\u003cstrong\\u003e2.2.2.7.\\u003c/strong\\u003e\\u003cem\\u003e3-(4-Bromophenyl)quinazoline-2,4(1H,3H)-dione\\u003c/em\\u003e \\u003cstrong\\u003e(4g)\\u003c/strong\\u003e\\u003c/h2\\u003e\\n \\u003cp\\u003eYield = 37.08%; m.p.= 240-243\\u003csup\\u003eo\\u003c/sup\\u003eC; IR(KBr): ν\\u003csub\\u003emax\\u003c/sub\\u003e = 3479 (NH Str, ArNH\\u003csub\\u003e2\\u003c/sub\\u003e), 2864 (C-H Str, Ar), 1725 (C = O, Str), 1690 (C = O Str, Cyclic CONH), 1452 (C = C Str, Ar), 1281 (C-N Str,Ar), 909.14 (NH, oop, wagging), 742 (C-H, oop, Ar), 680.5 (C-Br, Str) cm\\u003csup\\u003e− 1\\u003c/sup\\u003e, \\u003csup\\u003e1\\u003c/sup\\u003eH NMR (400MHz, DMSO-\\u003cem\\u003ed\\u003c/em\\u003e\\u003csub\\u003e\\u003cem\\u003e6\\u003c/em\\u003e\\u003c/sub\\u003e) \\u003cem\\u003eδ\\u003c/em\\u003e = 6.0 (s, 1H\\u003csub\\u003eamide\\u003c/sub\\u003e), 7.27 (t, 4H, ArH), 7.75 (s, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.13 (d, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e) ppm \\u003csup\\u003e13\\u003c/sup\\u003eC NMR (400MHz, DMSO-d\\u003csub\\u003e6\\u003c/sub\\u003e) 100MHz \\u003cem\\u003eδ\\u003c/em\\u003e = 118.7, 121.7, 123.8, 124.5, 127.7, 131.8, 132.4, 137.9, 150.0, 158.9 ppm; MS (m/z): 317.543 ([M\\u003csup\\u003e+\\u003c/sup\\u003e]), 306.811, 291.002, 278.053, 260.811, 242.443, 233.506, 224.552 C\\u003csub\\u003e14\\u003c/sub\\u003eH\\u003csub\\u003e9\\u003c/sub\\u003eN\\u003csub\\u003e2\\u003c/sub\\u003eO\\u003csub\\u003e2\\u003c/sub\\u003eBr (316.9)\\u003c/p\\u003e\\n \\u003c/div\\u003e\\n \\u003cdiv id=\\\"Sec14\\\"\\u003e\\n \\u003ch2\\u003e\\u003cstrong\\u003e2.2.2.8.\\u003c/strong\\u003e\\u003cem\\u003e3-(3-Nitrophenyl)quinazoline-2,4(1H,3H)-dione\\u003c/em\\u003e \\u003cstrong\\u003e(4h)\\u003c/strong\\u003e\\u003c/h2\\u003e\\n \\u003cp\\u003eYield = 79.49%; m.p.= 233-235\\u003csup\\u003eo\\u003c/sup\\u003eC; IR(KBr): ν\\u003csub\\u003emax\\u003c/sub\\u003e = 3484 (NH Str, ArNH\\u003csub\\u003e2\\u003c/sub\\u003e); 2897.5, 2869.2 (C-H Str, Ar); 1719.5 (C = O, Str); 1690 (C = O Str, Cyclic CONH); 1507.5, 1366.2 (N-O Str, NO\\u003csub\\u003e2\\u003c/sub\\u003e); 1455 (C = C Str, Ar); 1281.4 (C-N Str, Ar); 885.58 (N-H, oop, wagging); 744.23 (C-H, oop, Ar) cm\\u003csup\\u003e− 1\\u003c/sup\\u003e, \\u003csup\\u003e1\\u003c/sup\\u003eH NMR (400MHz, DMSO-\\u003cem\\u003ed\\u003c/em\\u003e\\u003csub\\u003e\\u003cem\\u003e6\\u003c/em\\u003e\\u003c/sub\\u003e) \\u003cem\\u003eδ\\u003c/em\\u003e = 6.0 (s, 1H\\u003csub\\u003eamide\\u003c/sub\\u003e), 7.0-7.6 (t, 4H, ArH), 7.75 (s, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.13 (d, 1H \\u003csub\\u003ebenzene\\u003c/sub\\u003e), 8.57 (s, 1H\\u003csub\\u003ebenzene\\u003c/sub\\u003e) ppm, \\u003csup\\u003e13\\u003c/sup\\u003eC NMR (400MHz, DMSO-d\\u003csub\\u003e6\\u003c/sub\\u003e) 100MHz \\u003cem\\u003eδ\\u003c/em\\u003e = 114.6, 119.5, 121.7, 123.3, 124.5, 127.7, 129.9, 132.4, 133.7, 137.9, 148.2, 150.0, 158.9 ppm; MS (m/z): 283.016 ([M\\u003csup\\u003e+\\u003c/sup\\u003e]), 278.883, 265.441, 251.222, 244.698, 238.446, 225.832 C\\u003csub\\u003e14\\u003c/sub\\u003eH\\u003csub\\u003e9\\u003c/sub\\u003eN\\u003csub\\u003e3\\u003c/sub\\u003eO\\u003csub\\u003e4\\u003c/sub\\u003e (283)\\u003c/p\\u003e\\n \\u003c/div\\u003e\\n \\u003c/div\\u003e\\n \\u003c/div\\u003e\\n\\u003c/div\\u003e\\n\\u003cdiv id=\\\"Sec15\\\"\\u003e\\n \\u003ch2\\u003e2.3. Molecular Docking\\u003c/h2\\u003e\\n \\u003cp\\u003eThe inhibitory activity of the 3-Substituted quinazoline-2,4(1H,3H)-dione derivatives with respect to antibacterial, antifungal, and anticonvulsant activities was analyzed using a molecular docking strategy. The protein structures of interest, identified by their Protein Data Bank (PDB) accession codes 7VPR, 6BPP, and 2Z5X [18–20] were obtained from the RCSB Protein Data Bank repository in PDB file format. The active sites of each protein structure were pre-determined by the construction of a grid box around the co-crystallized ligand. The protein molecules were prepared using AutoDock Tools [21, 22]. Restoring residues, eliminating water, introducing polar hydrogens, and assigning Kollman charges were all steps in this procedure. The protein structures were subsequently converted to pdbqt format for further analysis. Three-dimensional conformations of the ligand molecules were generated using the Merck Molecular Force Field 94 (MMFF94) implemented in Auto Dock Vina. These 3D ligand structures were then exported and converted to pdbqt format utilizing Open Babel GUI. A Perl script was executed to facilitate the molecular docking of all ligand molecules against the aforementioned protein structures. The binding affinities, expressed as docking scores, were calculated for each ligand-receptor interaction and reported in kcal/mol. Pymol and Discovery Studio Visualizer was further used to gain insights into the molecular interaction study of the ligands with the best binding affinity against each protein.\\u003c/p\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e2.4. Biological Evaluation\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e2.4. 1. Animals\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003eThe anticonvulsant efficacy of compounds 4(a-h) was assessed using the maximal electroshock seizure (MES) model, alongside an evaluation of their neurotoxicity profile. The study employed Wistar strain albino rats (100–250 g) randomly allocated into three groups (n = 6 per group): control, standard, and test. Prior to and throughout the experimental period, subjects were maintained on a standard laboratory pellet diet (Hindustan Lever Ltd., India) with ad libitum access to water. Following group assignment, animals were allowed a 2–3 day acclimatization period in the novel environment before commencement of the experimental protocol. All procedures adhered strictly to the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) guidelines for the anticonvulsant screening of synthesized compounds. The study was reviewed and approval for the conducting of activity was given by the Institutional Animal Ethics Committee (IAEC Registration No-838/PO/Re/S/04/CPCSEA), Dr. K.N. Modi Institute of Pharmaceutical Education and Research, Modinagar, Uttar Pradesh, India.\\u003c/p\\u003e\\n \\u003cdiv id=\\\"Sec16\\\"\\u003e\\n \\u003ch2\\u003e2.4.2. Anticonvulsant Activity\\u003c/h2\\u003e\\n \\u003cdiv id=\\\"Sec17\\\"\\u003e\\n \\u003ch2\\u003e2.4.2.1. Maximal electroshock seizure (MES) method\\u003c/h2\\u003e\\n \\u003cp\\u003eAll the synthesized compounds were subjected to MES method for anticonvulsant activity. Seizures were induced in all three groups by delivering electroshock of 60 Hz of alternating current 150mA for 0.2 sec by means of ear clip electrodes using electro-convulsiometer. 1% carboxymethylcellulose sodium (CMC) solution was used as control; phenytoin sodium (30mg/kg) was used as standard drug and administered through intraperitoneal route prior to evaluation. The dosage (30, 60, and 100 mg/kg body weight) were prepared as suspensions of synthesized compounds in 1% carboxymethylcellulose (CMC) and administered through intraperitoneal (i.p.) injection. Assessments were conducted at 0.5 and 4 hours post-administration. The anticonvulsant activity was quantified by observing the absence or reduction of the hind limb tonic extensor phase, a characteristic indicator of seizure severity in this model [13, 16, 23].\\u003c/p\\u003e\\n \\u003c/div\\u003e\\n \\u003cdiv id=\\\"Sec18\\\"\\u003e\\n \\u003ch2\\u003e2.4.2.2. Neurotoxicity Screening (Rotarod Method)\\u003c/h2\\u003e\\n \\u003cp\\u003eMotor function assessment was conducted using the rotarod test to evaluate minimal motor impairment in the subjects. The apparatus consisted of a rotating rod with a diameter of 3.2 cm, initially set to a speed of 6 rpm with acceleration capability. Subjects were habituated to the device prior to testing. Following the training period, test compounds were administered intraperitoneally at a dose of 100 mg/kg body weight. Neurotoxicity was defined as the lack of ability to uphold balance on the rotating rod for atleast 60 seconds in each of four consecutive experiments. This method provided a quantitative measure of motor coordination and balance under the influence of the administered compounds [13, 16, 23].\\u003c/p\\u003e\\n \\u003c/div\\u003e\\n \\u003c/div\\u003e\\n \\u003cdiv id=\\\"Sec19\\\"\\u003e\\n \\u003ch2\\u003e2.4.3 Antibacterial and Antifungal Activity\\u003c/h2\\u003e\\n \\u003cp\\u003eThe antimicrobial efficacy against bacterial and fungal pathogens was evaluated through in vitro assays using pure cultures. In vitro susceptibility testing was conducted employing the following methodologies:\\u003c/p\\u003e\\n \\u003cdiv id=\\\"Sec20\\\"\\u003e\\n \\u003ch2\\u003e2.4.3.1. Tube Dilution Method\\u003c/h2\\u003e\\n \\u003cp\\u003eDilutions of the synthesized compounds were prepared in growth medium to encompass their clinically relevant concentration ranges. Each tube, including a control without antimicrobial agent, was inoculated with an equivalent volume of broth containing 105–106 CFU/mL of bacteria. Following overnight incubation, the tubes were assessed for visible turbidity. Tube dilution method was resulted in the determination of antibacterial susceptibility in liquid media [24], and MIC determination for select derivatives.\\u003c/p\\u003e\\n \\u003c/div\\u003e\\n \\u003cdiv id=\\\"Sec21\\\"\\u003e\\n \\u003ch2\\u003e2.4.3.2. Agar Diffusion Method\\u003c/h2\\u003e\\n \\u003cp\\u003ePetri dishes containing agar medium were prepared using the pour plate method. The medium was subsequently inoculated with the target microorganisms. For the agar dilution assay, varying concentrations of antibiotics were incorporated into the agar medium to assess both aerobic and anaerobic microbial growth and incubated at 37°C for 24 hours. The antimicrobial agents diffused through the agar, resulting in clear zones of inhibition. The diameters of these zones were measured to estimate the efficacy of the antimicrobial substances [24]. Nutrient agar media was used for the purpose of preparation of agar plates for the growth of selected bacteria namely Gram positive strains- \\u003cem\\u003eB. subtilis\\u003c/em\\u003e and \\u003cem\\u003eS. aureus\\u003c/em\\u003e and Gram negative strains- \\u003cem\\u003eE. coli\\u003c/em\\u003e and \\u003cem\\u003eP. aeruginosa\\u003c/em\\u003e. Whereas Sabouraud’s agar media was used for the preparation of petri plates meant for the growth of fungi namely \\u003cem\\u003eA. niger\\u003c/em\\u003e and \\u003cem\\u003eC. albicans\\u003c/em\\u003e [25]. The Agar well diffusion test was performed by using nutrient agar medium as per the procedure given by Venkatesarlu \\u003cem\\u003eet al.\\u003c/em\\u003e [26]. Each of the agar media-nutrient and Sabrouraud's was autoclaved for 15 minutes at 121ºC to ensure sterilityand then they were removed from the water bath immediately to cool them at 45ºC-55ºC [26]. Sterilization of petri dishes and pipettes was conducted in an oven at 150°C for 60 minutes. Subsequently, 25 mL of molten agar medium was aseptically dispensed into each sterilized Petri dish (10 cm diameter). To prepare inocula, bacterial cultures were propagated in 80 mL of nutrient broth, while fungal cultures were grown on Sabouraud's agar media. Both were incubated in 250 mL Erlenmeyer flasks. The inoculum flasks were incubated at 37ºC in case of bacteria for 18 hours and at 25ºC in case of fungi for 25 hours. Approximately 0.5 mL of inoculum broth containing various bacterial and fungal strains was introduced into separate petri dishes. A rotational technique was employed to ensure uniform distribution of the contents. Subsequently, the inoculated medium was allowed to solidify at ambient temperature. Standard drug used for antibacterial evaluation was ciprofloxacin and its stock solution was prepared using dissolving 1mg of drug in 1ml of dimethylsulfoxide (DMSO) to obtain 1000µg/mL of solution which was then serially diluted to obtain solutions of concentration 25, 50, 75, 100µg/mL. Griseofulvin was used as standard for the antifungal activity and its stock solution was prepared by dissolving 1mg of drug in 1mL of DMSO to obtain 1000µg/mL of solution which was then serially diluted to obtain solutions of concentration 25, 50, 75, 100µg/mL. All the test compounds were prepared by dissolving 1mg of drug in 1mL of DMSO to obtain 1000 µg/mL of solution which was then serially diluted to obtain solutions of concentration 25, 50, 75, 100 µg/mL [27]. 0.1 mL of the fraction of standard drug and test compounds were added in the wells and labeled accordingly. Petri-dishes were then kept in cool place for one hour to allow diffusion of solution into the medium and further incubated at 37ºC for 18 hours in case of bacteria and at 25ºC for 25 hours in case of fungi. Inhibition zones were observed surrounding the wells, and their diameters were measured in millimeters [26].\\u003c/p\\u003e\\n \\u003c/div\\u003e\\n \\u003c/div\\u003e\\n\\u003c/div\\u003e\"},{\"header\":\"3. Results and Discussion\",\"content\":\"\\u003cp\\u003eThe compounds \\u003cb\\u003e3(a-h)\\u003c/b\\u003e and \\u003cb\\u003e4(a-h)\\u003c/b\\u003e were synthesized as per reactions given in the \\u003cb\\u003escheme 1\\u003c/b\\u003e and \\u003cb\\u003escheme 2\\u003c/b\\u003e respectively.\\u003c/p\\u003e \\u003cdiv id=\\\"Sec23\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.1. Molecular Docking\\u003c/h2\\u003e \\u003cp\\u003eTo figure out the molecular mechanism behind the antibacterial activity of the 3-Substituted quinazoline-2,4 (1H,3H)-dione derivatives, a molecular docking approach was conducted against the Lipid A export ATP-binding/permease protein MsbA of gram-negative bacteria (PDB ID: 6BPP). Among the eight derivatives, compounds 4d and 4h showed the highest binding affinity i.e. -8.7 kcal/mol against MsbA protein. Compound 4d showed significant molecular interactions by demonstration of alkyl and pi-alkyl interaction at LEU171, LEU263, MET291, VAL178, ALA262, LEU294, pi-sigma interactions at LEU298 and ILE182, along with a halogen bond formation at ALA259.\\u003c/p\\u003e \\u003cp\\u003eOn the other hand compound 4h demonstrated pi-alkyl interactions at VAL178, ALA262, ALA259, and LEU171, along with pi-sigma bond formation at LEU298 and ILE182 (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e, Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eTo assess the antifungal activity of the synthesized compounds, a molecular docking was conducted against the fungal sterol uptake control protein 2 (Upc2) (PDB ID: 7VPR). Compound 4a exhibited highest binding affinity (-10 kcal/mol) against Upc2 protein. The significant molecular interactions of compound 4a were observed by demonstration of hydrogen bond formation at MET858, alkyl and pi-alkyl interactions at PRO836, VAL750, MET901, ARG859, and LEU906, pi-pi stacked bond formation at PHE905, and pi-sigma bond formation at VAL890 (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e, Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eIn-silico screening was also conducted against Human Monoamine Oxidase A (MAO-A) (PDB ID: 2Z5X) to gain insights of mechanism behind the anticonvulsant activity of aforesaid derivatives. Among all, compound 4h exhibited highest binding affinity of -10.7 kcal/mol against MAO-A protein. The significant interactions of compound 4h were demonstrated by hydrogen bond formation at TYR444, TYR69, ALA68, MET445, pi-donor hydrogen bond formation at GLY67 and TYR407, pi-pi stacked and pi-pi t-shaped bond formation at TYR444 and TYR407, along with pi-alkyl bond formation at ILE335 (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003e, Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\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\\u003eBinding affinity scores of all the 3-Substituted quinazoline-2,4 (1H,3H)-dione derivatives against three different protein structures\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"4\\\"\\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=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eCompound\\u003c/p\\u003e \\u003cp\\u003eName\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eBinding affinity against PDB ID:6BPP (kcal/mol)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eBinding affinity against PDB ID:7VPR (kcal/mol)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eBinding affinity against PDB ID:2Z5X (kcal/mol)\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4a\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e-8.4\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-9.6\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4b\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e-8.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-9.6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-9.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4c\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e-8.2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-9.6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-9.3\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4d\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e-8.7\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-9.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-10.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e-8.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-9.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-10.2\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4f\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e-6.8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-8.4\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-8.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4g\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e-8.4\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-9.8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-9.6\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4h\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e-8.7\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-9.6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-10.7\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec24\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.2. Anticonvulsant Activity\\u003c/h2\\u003e \\u003cp\\u003eThe synthesized compounds 4 (a-h) demonstrated anticonvulsant activities which were evident through reduction in the duration of the hind limb tonic extensor phase (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e). Among all the compounds synthesized, compound \\u003cb\\u003e4h\\u003c/b\\u003e was found to be equipotent to the drug phenytoin sodium as it abolished seizures at the dose 30 mg/Kg at 0.5h and after 4h of administration. Compound 4d demonstrated anticonvulsant activity at a dose of 60 mg/kg, with seizure inhibition observed at both 0.5 and 4 hours post-administration.Compound \\u003cb\\u003e4a\\u003c/b\\u003e showed inhibited seizures at 60mg/kg at 0.5h and at 100mg/kg, after 4h of administration. Whereas compounds \\u003cb\\u003e4b\\u003c/b\\u003e, \\u003cb\\u003e4c\\u003c/b\\u003e, \\u003cb\\u003e4e\\u003c/b\\u003e, \\u003cb\\u003e4f\\u003c/b\\u003e and \\u003cb\\u003e4g\\u003c/b\\u003e inhibited seizures at 100mg/kg at 0.5h and after 4.0h of administration. Neurotoxicity screening was conducted on compounds 4(a-h) in which none of the synthesized compounds exhibited neurotoxic effects at a dose of 100 mg/kg when evaluated at 0.5 and 4 hours post-administration (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\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\\u003eAnticonvulsant activity of the synthesized compounds in the maximal electroshock seizure method\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"3\\\"\\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 \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eCompound\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c3\\\" namest=\\\"c2\\\"\\u003e \\u003cp\\u003eMES\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e0.5h\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4.0h\\u003c/b\\u003e\\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\\u003e4a\\u003c/b\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u003cb\\u003e4b\\u003c/b\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u003cb\\u003e4c\\u003c/b\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u003cb\\u003e4d\\u003c/b\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u003cb\\u003e4e\\u003c/b\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u003cb\\u003e4f\\u003c/b\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u003cb\\u003e4g\\u003c/b\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u003cb\\u003e4h\\u003c/b\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u003cb\\u003ePhenytoin\\u003c/b\\u003e\\u003csup\\u003e\\u003cb\\u003eb\\u003c/b\\u003e\\u003c/sup\\u003e \\u003cb\\u003eSodium\\u003c/b\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u003cb\\u003eControl 1% (w/v)CMC\\u003c/b\\u003e\\u003csup\\u003e\\u003cb\\u003ec\\u003c/b\\u003e\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e60\\u003c/p\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003cp\\u003e60\\u003c/p\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003cp\\u003e60\\u003c/p\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003cp\\u003e1\\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\\u003e \\u003csup\\u003e \\u003cb\\u003ea\\u003c/b\\u003e \\u003c/sup\\u003eIntraperitoneal (i.p.) injections of the compound were administered to rats at doses of 30, 60, and 100 mg/kg. The values presented in the table represent the minimum dose at which bioactivity was observed in 50% or more of the test subjects. Examinations were conducted at 0.5 and 4.0 hours post-administration.\\u003c/p\\u003e \\u003cp\\u003e \\u003csup\\u003e \\u003cb\\u003eb\\u003c/b\\u003e \\u003c/sup\\u003eReference drug phenytoin sodium (30mg/Kg).\\u003c/p\\u003e \\u003cp\\u003e \\u003csup\\u003e \\u003cb\\u003ec\\u003c/b\\u003e \\u003c/sup\\u003eControl compound 1% (w/v) CMC solution.\\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\\u003eEvaluation of neurotoxic potential in the synthesized novel compounds\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"3\\\"\\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 \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eCompound\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c3\\\" namest=\\\"c2\\\"\\u003e \\u003cp\\u003eNeurotoxicity Screening\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0.5h\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e4.0h\\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\\u003e4a\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4b\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4c\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4d\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4e\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4f\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4g\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4h\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003ePhenytoin\\u003c/b\\u003e\\u003csup\\u003e\\u003cb\\u003eb\\u003c/b\\u003e\\u003c/sup\\u003e \\u003cb\\u003eSodium\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0/6\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"3\\\"\\u003e\\u003csup\\u003ea\\u003c/sup\\u003eDose of 100 mg/kg were administered through i.p injection in rat. Values indicate number of animals that showed neurotoxicity out of six animals\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec25\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.3. Antibacterial activity\\u003c/h2\\u003e \\u003cp\\u003eThe in vitro antibacterial efficacy of novel compounds 4(a-h) was evaluated using the agar diffusion method. The assay was conducted against both Gram-positive (B. subtilis and S. aureus) and Gram-negative (E. coli and P. aeruginosa) bacterial strains. All compounds 4(a-h) were assessed for their antibacterial activity at a concentration of 100 \\u0026micro;g/mL.The zone of inhibition (mm) was determined for each compound as measure of their activity along with Ciprofloxacin as standard drugs (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e). Outperforming all the other synthesised compounds, 4f showed strongest efficacy against all the selected bacteria, preventing their development at a concentration of 100 \\u0026micro;g/mL (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig7\\\" class=\\\"InternalRef\\\"\\u003e7\\u003c/span\\u003e). However, all the synthesized compounds inhibited the growth of selected bacteria.\\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 4\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eIn vitro\\u003c/em\\u003e antibacterial activity of compounds 4(a-h) against selected strains\\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\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eCompound\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"4\\\" nameend=\\\"c5\\\" namest=\\\"c2\\\"\\u003e \\u003cp\\u003eZone of Inhibition\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eS. aureus\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eB\\u003c/em\\u003e. \\u003cem\\u003esubtilis\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eE\\u003c/em\\u003e. \\u003cem\\u003ecoli\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eP. aeruginosa\\u003c/em\\u003e\\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\\u003e4a\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.8 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e6.8 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e7.1 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8.0 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4b\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e9 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e11 (25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e5 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e7 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4c\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e6 (25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e8 (12.5)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e7 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8 (25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4d\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.2 (25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5.9 (25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.3 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8.7 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4e\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e8.5 (25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e8.6 (25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e9.1(12.5)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e9.4(12.5)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4f\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e12.1(6.25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e13.5(6.25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e11.8(12.5)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e11.5 (12.5)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4g\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e6.6 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e6.8(25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e6.2(25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e7(6.25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4h\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e8.1(25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e8.9(50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e7.5(100)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e7.9(100)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eCiprofloxacin\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e18(12.5)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e19(6.25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e19(12.5)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e17(6.25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"5\\\"\\u003eValues in bracket are MIC values (\\u0026micro;g/mL)\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec26\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.4. Antifungal activity\\u003c/h2\\u003e \\u003cp\\u003eThe antifungal susceptibility of the developed compounds was evaluated using the agar diffusion method against the target bacteria \\u003cem\\u003eviz\\u003c/em\\u003e. Fungus strains \\u003cem\\u003eC. albicans\\u003c/em\\u003e and \\u003cem\\u003eA. niger\\u003c/em\\u003e. The antifungal susceptibility of all synthesized compounds was evaluated at a concentration of 100 \\u0026micro;g/mL using the agar diffusion method.The zone of inhibition (in mm) was determined for each compound as measure of their activity along with griseofulvin as standard drugs (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e). Among all the compounds synthesized, compound \\u003cb\\u003e4b\\u003c/b\\u003e showed maximum activity against selected fungal strains and it showed activity greater than the standard drug (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig8\\\" class=\\\"InternalRef\\\"\\u003e8\\u003c/span\\u003e). Compound \\u003cb\\u003e4d\\u003c/b\\u003e also showed greater activity but was less active than the standard drug.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab5\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 5\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eIn vitro\\u003c/em\\u003e antifungal activities of compounds 4(a-h) against selected strains\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"3\\\"\\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 \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eCompound\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c3\\\" namest=\\\"c2\\\"\\u003e \\u003cp\\u003eZone of Inhibition\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eC.albicans\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eA.niger\\u003c/em\\u003e\\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\\u003e4a\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e12 (100)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e13 (100)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4b\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e25 (6.25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e25 (6.25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4c\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e12 (100)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e13 (100)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4d\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e21(25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e22 (6.25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4e\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e19 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e18 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4f\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e18 (25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e17 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4g\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e19 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e19 (50)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4h\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e16 (100)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e18 (100)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eGriseofulvin\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e23 (6.25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e23 (6.25)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"3\\\"\\u003eValues in bracket are MIC values (\\u0026micro;g/mL)\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"4. Conclusion\",\"content\":\"\\u003cp\\u003eThe main objective of this research was to synthesized a series of quinazoline-2,4-dione derivatives with medicinal activity and minimal adverse effects.The therapeutic potential of 3-substituted quinazoline-2,4(1H,3H)-diones derivatives may be the new vista of research there in this particular area.A series of 3-substituted quinazoline2,4(1H,3H)-dione derivatives was prepared starting from reaction of isatoic anhydride with different aromatic amines by taking dimethylformamide as solvent to yield 2-amino-N-benzamide derivatives in first scheme followed by the reaction with triphosgene in the presence of potassium carbonate by taking tetrahydrofuran to bring about cyclization to finally produce 3-substituted-quinazoline-2,4(1H,3H)-dione derivatives in second scheme. All the synthesized compounds were purified by re-crystallization and identified on the basis of physicochemical characterization melting point and spectral characterization by FTIR, H\\u003csup\\u003e1\\u003c/sup\\u003eNMR, \\u003csup\\u003e13\\u003c/sup\\u003eCNMR and MALDI-MS spectral method. In vitro evaluation using the MES method revealed significant anticonvulsant activity of the synthesized compounds compared to the standard drug, phenytoin sodium. However, all of the synthesized compounds also exhibited antibacterial and anti-fungal activities when compared with the standard drugs- Ciprofloxacin and Griseofulvin for antimicrobial screening purpose. Compound \\u003cb\\u003e4h\\u003c/b\\u003e was found to be equipotent to the drug phenytoin sodium which makes us conclude that it may serve as lead for the synthesis of new compounds with anticonvulsant action. Compound \\u003cb\\u003e4f\\u003c/b\\u003e was effective against all the selected bacteria and inhibited their growth so it can be used further for the synthesis of compounds with great antibacterial activity. Compound \\u003cb\\u003e4b\\u003c/b\\u003e exhibited activity greater than the standard drug griseofulvin therefore; it may serve as lead for the synthesis of series of new compounds with greater antifungal activity.\\u003c/p\\u003e\"},{\"header\":\"Abbreviations\",\"content\":\"\\u003cp\\u003e\\u003csup\\u003e1\\u003c/sup\\u003eH NMR; Proton Nuclear Magnetic Resonance, AMPA; \\u0026alpha;-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid, \\u003cem\\u003eA.\\u0026nbsp;\\u003c/em\\u003e\\u003cem\\u003eniger; Aspergillus niger,\\u0026nbsp;\\u003c/em\\u003eATP; Adenosine Triphosphate, \\u003cem\\u003eB. Subtilis;\\u0026nbsp;\\u003c/em\\u003e\\u003cem\\u003eBacillus subtilis,\\u0026nbsp;\\u003c/em\\u003e\\u003cem\\u003eC.\\u0026nbsp;\\u003c/em\\u003e\\u003cem\\u003ealbicans; Candida albicans,\\u0026nbsp;\\u003c/em\\u003eDMEM; Dulbecco\\u0026apos;s Modified Eagle\\u0026apos;s Media, DMF; Dimethylformamide, DMSO; Dimethyl Sulfoxide, DNA; Deoxyribonucleic Acid, \\u003cem\\u003eE. coli\\u003c/em\\u003e; \\u003cem\\u003eEscherichia coli,\\u0026nbsp;\\u003c/em\\u003eERG; Electron Releasing Group, EWG; Electron-Withdrawing Group, FT-IR; Fourier-Transform Infrared Spectroscopy, IC\\u003csub\\u003e50\\u003c/sub\\u003e; Half-Maximal Inhibitory Concentration, HCl; Hydrochloric Acid, m; Meta, o; Ortho, p; Para, PBS; Phosphate-Buffered Saline, PCT; Proximal Convoluted Tubule, \\u003cem\\u003eP.\\u0026nbsp;\\u003c/em\\u003e\\u003cem\\u003eaeruginosa; Pseudomonas aeruginosa,\\u0026nbsp;\\u003c/em\\u003eppm; parts per million, \\u003cem\\u003e\\u0026nbsp;\\u003c/em\\u003eRBF; Round Bottom Flask, SAR; Structure-activity Relationship, \\u003cem\\u003eS. aureus\\u003c/em\\u003e; \\u003cem\\u003eStaphylococcus aureus\\u003c/em\\u003e, THF; Tetrahydrofuran, TLC; Thin-Layer Chromatography, TPSA; Total Polar Surface Area\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgement\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe synthetic work and biological activity supported by Dr. KNMIPER, Modinagar, Uttar Pradesh, India is greatly acknowledged. Authors duly acknowledge Kusuma School of Biological Science, Indian Institute of Technology (IIT), Delhi, India for providing MALDI-MS analysis. Authors are also thankful to Sophisticated Analytical Instrument Facility (SAIF), Panjab University, Chandigarh, India for providing spectroscopic facilities.\\u0026nbsp;\\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 and its Supplementary Information files. Should any raw data files be needed in another format they are available from the corresponding author upon reasonable request. Source data are provided with this paper.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthor Contributions\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eConceptualization and study design were carried out by the corresponding author. Chemical synthesis and characterization of the compounds were performed by Neelam Sharma. Biological evaluation, including anticonvulsant, antibacterial, and antifungal studies, was conducted by Manish Kumar and Vijay Kumar Sharma. Molecular docking studies and computational analysis were performed by Ratul Bhowmik and Corresponding author. Data analysis and interpretation were carried out by Manish Kumar. The manuscript was written by Neelam Sharma and corresponding author and reviewed and approved by all authors.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConflict of Interests\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors report no conflict of interests.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\u003cli\\u003e\\u003cspan\\u003eLi X, Lee YR. Facile One-Pot Synthesis of Quinazoline-2,4-dione Derivatives and Application to Naturally Occurring Alkaloids from Zanthoxylum Arborescens. Bulletin of the Korean Chemical Society. 2011;32:2121\\u0026ndash;2124\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eJin HZ, Du JL, Zhang WD, Yan SK, Chen HS, Lee JH, Lee JJ. A new quinazolinedione alkaloid from the fruits of Evodia officinalis. Fitoterapia. 2008;79:317\\u0026ndash;318\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMichael, JP. Quinoline, quinazoline and acridone alkaloids. Nat. Prod. Rep. 2001;18: 543\\u0026ndash;559\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eChawlaA,BatraC. Recent advances of quinazolinone derivatives as marker for various biological activities. Int. Res. J. Pharm. 2013;4:49\\u0026ndash;58\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eT Panneerselvam, Kumar PV. Quinazoline Marketed drugs- A Review.Research in Pharmacy. 2011;1: 1\\u0026ndash;21\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eKaur H, Kumar B, Medhi B. Antiepileptic drugs in development pipeline: A recent update. eNeurologicalSci.2016;4:42\\u0026ndash;51\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eDadiboyena S,Arfaoui A, Amri H, Piedrafita FJ, Nefzi A. Diversity oriented synthesis and IKK inhibition of aminobenzimidazole tethered quinazoline-2,4-diones, thioxoquinazolin-4-ones, benzodiazepine-2,3,5-triones, isoxazoles and isoxazolines. Bioorg. Med. Chem. Lett. 2015;25:685\\u0026ndash;689\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLanglois M, Soulier JL, Rampillon V, Gallais C, Br\\u0026eacute;mont B, Shen S, Yang D, Giudice A, Sureau F. Synthesis of quinazoline-2,4-dione and naphthalimide derivatives as new 5-HT3 receptor antagonists. Eur. J. Med. Chem. 1994;29:925\\u0026ndash;940\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eDangi RR, Chundawat NS, Talesara GL.A convenient synthesis of Ethoxyphthalimide derivatized quinazoline assembled pyrimidine and pyridine via common intermediate chalcone and their antimicrobial agents.World J. Pharm. Res.2015;4:1400\\u0026ndash;1413\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eNencini A, Pratelli C, Quinn JM, Salerno M, Tunici P, De Robertis A, Valensin S, Mennillo F, Rossi M, Bakker A, Benicchi T, Cappelli F, Turlizzi E, Nibbio M, Caradonna NP, Zanelli U, Andreini M, Magnani M, Varrone M. Structure\\u0026ndash;activity relationship and properties optimization of a series of quinazoline-2,4-diones as inhibitors of the canonical wnt pathway. Eur. J. Med. Chem. 2015;95:526\\u0026ndash;545\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eYao H, Ji M, Zhu Z, Zhou J, Cao R, Chen X, Xu B. Discovery of 1-substituted benzyl-quinazoline-2,4(1H,3H)-dione derivatives as novel poly(ADP-ribose)polymerase-1 inhibitors. Bioorg Med Chem. 2015;23:681\\u0026ndash;693\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eWhite HS, Smith MD, Wilcox KS. Mechanisms of action of antiepileptic drugs. Int Rev Neurobiol. 2007;81:85\\u0026ndash;110\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003ePrashanth MK, Madaiah M, Revanasiddappa HD, Veeresh B. Synthesis, anticonvulsant, antioxidant and binding interaction of novel \\u003cem\\u003eN\\u003c/em\\u003e-substituted methylquinazoline-2,4(1H, 3H)-dione derivatives to bovine serum albumin: A structure\\u0026ndash;activity relationship study.Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy.2013;110:324\\u0026ndash;332\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSiddiqui N, Ahsan W, Alam MS, Ali R, Srivastava K, AhmedS.Anticonvulsant activity of a combined pharmacophore of pyrazolo-pyridines with Lesser toxicity in mice. Bulletin of the Korean Chemical Society. 2011;32:576\\u0026ndash;582\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eVerma M, Pandeya SN, Singh KN, Stables JP. Anticonvulsant Activity of Schiff Bases of Isatin Derivatives. Acta Pharmaceutica.2004;54:49\\u0026ndash;56\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eKumar D, Sharma VK, Kumar R, Singh T, Singh H, Singh AD, Roy, R. Design, synthesis and anticonvulsant activity of some new 5,7-dibromoisatin semicarbazone derivatives. EXCLI Journal. 2013;12:628\\u0026ndash;640\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMabkhot YN, Al-Majid AM, Barakat A, Al-Showiman SS, Al-Har MS, Radi S, Naseer MM, Hadda TB. Synthesis and Biological Evaluation of 2-Aminobenzamide Derivatives as Antimicrobial Agents: Opening/Closing Pharmacophore Site. International Journal of Molecular Sciences. 2014;15:5115\\u0026ndash;5127\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eElalouf A. \\u003cem\\u003eIn-silico\\u003c/em\\u003e Structural Modeling of Human Immunodeficiency Virus Proteins. Biomed Eng Comput Biol. 2023;14:11795972231154402\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eHo H, Miu A, Alexander MK, Garcia NK, Oh A, Zilberleyb I, Reichelt M, Austin CD, Tam C, Shriver S, Hu H, Labadie SS, Liang J, Wang L, Wang J, Lu Y, PurkeyHE, Quinn J, Franke Y, Clark K, Beresini MH, Tan MW, Sellers BD, Maurer T, Koehler MFT, Wecksler AT, Kiefer JR, Verma V, Xu Y, Nishiyama M, Payandeh J, Koth CM. Structural basis for dual-mode inhibition of the ABC transporter MsbA. Nature. 2018;557:196\\u0026ndash;201\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSon SY, Ma J, Kondou Y, Yoshimura M, Yamashita E, Tsukihara T. Structure of human monoamine oxidase A at 2.2-A resolution: the control of opening the entry for substrates/inhibitors. Proc Natl Acad Sci U S A. 2008;105:5739\\u0026ndash;44\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eEberhardtJ, Santos-Martins D, Tillack AF, Forli S. AutoDock Vina 1.2.0: New Docking Methods, Expanded Force Field, and Python Bindings. J. Chem. Inf. Model. 2021;61:3891\\u0026ndash;3898\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eTanchuk VY, Tanin VO, Vovk AI, Poda, G. A New, improved hybrid scoring function for Molecular Docking and scoring based on AutoDock and AutoDock Vina. Chem Biol Drug Des. 2016;87:618\\u0026ndash;625\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eKulkarniSK. Handbook of Experimental Pharmacology (3rd ed), Vallabh Prakashan, Delhi,1999;Reprint 2009:131\\u0026ndash;134\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eIndian Pharmacopoeia commission, Govt. of India. Micrbiological Assay of Antibiotics, Indian Pharmacopoeia, 2014;3(7):38\\u0026ndash;50\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eBlack WD. A comparison of several media types and basic techniques used to assess outdoor airborne fungi in Melbourne, Australia. PLoS One. 2020;15:e0238901\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eVenkatesarulu V, Kokate C,Rambhau D, Ciddi V. Antimicrobial activity of chemoconstituents of roots of salaciamacrosperma. Ancient science of life. 1992;12: 251\\u0026ndash;256\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eZaranappa, Vagdevi HM, Jayanna ND, Latha KP. Synthesis, Characterization and evaluation of Antibacterial Activity of some 3-Substitutedphenylquinazoline \\u0026ndash;\\u0026thinsp;2,4-diones. Der Pharma Chem. 2012;4: 1754\\u0026ndash;1758\\u003c/span\\u003e\\u003c/li\\u003e\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":true,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":false,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"researchsquare\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":true,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"/submission\",\"title\":\"Research Square\",\"twitterHandle\":\"researchsquare\",\"acdcEnabled\":true,\"dfaEnabled\":false,\"editorialSystem\":\"\",\"reportingPortfolio\":\"\",\"inReviewEnabled\":false,\"inReviewRevisionsEnabled\":true},\"keywords\":\"Quinazoline-2,4-dione, Heterocyclic scaffold, Molecular docking, Anticonvulsant activity, Antimicrobial agents\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-8566641/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-8566641/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eA series of novel 3-substituted quinazoline-2,4(1H,3H)-dione derivatives was designed, synthesized, and evaluated for their anticonvulsant, antibacterial, and antifungal activities. Anticonvulsant efficacy was assessed \\u003cem\\u003ein vivo\\u003c/em\\u003e using the maximal electroshock (MES) seizure model, while antibacterial and antifungal activities were screened against selected Gram-positive, Gram-negative bacterial, and fungal strains. To support the experimental findings and elucidate possible binding interactions, molecular docking studies were carried out against relevant biological targets for antibacterial (PDB ID: 6BPP), antifungal (PDB ID: 7VPR), and anticonvulsant activity (PDB ID: 2Z5X). Among the synthesized compounds, compound 4h exhibited significant anticonvulsant activity by effectively inhibiting extensor seizures in the MES model at a dose of 30 mg/kg, showing comparable efficacy to the reference drug phenytoin sodium. Compound 4f demonstrated broad-spectrum antibacterial activity against Gram-positive bacteria (\\u003cem\\u003eBacillus subtilis\\u003c/em\\u003e, \\u003cem\\u003eStaphylococcus aureus\\u003c/em\\u003e) and Gram-negative bacteria (\\u003cem\\u003eEscherichia coli\\u003c/em\\u003e, \\u003cem\\u003ePseudomonas aeruginosa\\u003c/em\\u003e), inhibiting microbial growth at 100 \\u0026micro;g/mL. Additionally, compound 4b displayed potent antifungal activity against \\u003cem\\u003eCandida albicans\\u003c/em\\u003e and \\u003cem\\u003eAspergillus niger\\u003c/em\\u003e at 100 \\u0026micro;g/mL, surpassing the activity of the standard antifungal agent griseofulvin. The study highlights the quinazoline-2,4-dione heterocyclic scaffold as a promising pharmacophore for the development of multifunctional bioactive agents with potential therapeutic relevance and reduced adverse effects. The combined \\u003cem\\u003ein silico\\u003c/em\\u003e and experimental findings provide a valuable foundation for further structural optimization and pharmacological exploration.\\u003c/p\\u003e\",\"manuscriptTitle\":\"From In Silico Design to Bioactivity: Synthesis and Pharmacological Profiling of 3-Substituted Quinazoline-2,4-dione Heterocycles\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2026-02-22 15:49:33\",\"doi\":\"10.21203/rs.3.rs-8566641/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"researchsquare\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":true,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"/submission\",\"title\":\"Research Square\",\"twitterHandle\":\"researchsquare\",\"acdcEnabled\":true,\"dfaEnabled\":false,\"editorialSystem\":\"\",\"reportingPortfolio\":\"\",\"inReviewEnabled\":false,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"c6bd7d15-24a5-4514-bd0b-93526863f128\",\"owner\":[],\"postedDate\":\"February 22nd, 2026\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"posted\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2026-04-21T16:25:17+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2026-02-22 15:49:33\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-8566641\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-8566641\",\"identity\":\"rs-8566641\",\"version\":[\"v1\"]},\"buildId\":\"XKTyCvWXoU3ODBz1xrDgd\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}