Towards new bioactive fluorine-containing 1,3,4-oxadiazole-amide derivatives: Synthesis, antibacterial activity, molecular docking and molecular dynamics simulation study

Towards new bioactive fluorine-containing 1,3,4-oxadiazole-amide derivatives: Synthesis, antibacterial activity, molecular docking and molecular dynamics simulation study | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Towards new bioactive fluorine-containing 1,3,4-oxadiazole-amide derivatives: Synthesis, antibacterial activity, molecular docking and molecular dynamics simulation study Fei Xiong, Yanjun Zhang, Jinlong Jiao, Yiren Zhu, Tianlu Mo, Yeji Li This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4148372/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 20 Jun, 2024 Read the published version in Molecular Diversity → Version 1 posted 9 You are reading this latest preprint version Abstract Through the approach of molecular hybridization, this study rationally designed and synthesized new trifluoromethyl-1,3,4-oxadiazole amide derivatives, denoted as 1a-1n . The findings reveal that these novel molecules exhibit potent inhibitory effects against various bacterial strains. Thereinto, compounds 1c , 1d , 1i , 1j and 1n , demonstrate relatively superior antimicrobial performance against B. cereus FM314, with a minimum inhibitory concentration (MIC) of 0.03907 µg/mL. Molecular docking analysis suggests the potential importance of the Ser57 and Thr125 amino acid residues in contributing to the inhibitory activity against B. cereus . The consistency of these results was further corroborated through subsequent molecular dynamics simulations and MMPBSA validations. The insights gained from this study serve to facilitate the rational design and efficient development of novel eco-friendly antimicrobial inhibitors based on the trifluoromethyl-1,3,4-oxadiazole amide scaffold. Bacillus cereus 1 3 4-Oxadiazole Synthesis Biological assays MD Simulation Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Bacterial diseases stemming from escalating antimicrobial resistance have emerged as a paramount concern in the public health domain in recent years [ 1 ]. The ubiquitous foodborne pathogen- Bacillus cereus , which contaminates a range of edible products, including flour-based items and dairy products, poses a significant health threat to both adults and children [ 2 ]. Research has elucidated that this bacterial species is ubiquitous in diverse environments, such as soil, water, air, and food matrices, existing in various forms, including vegetative cells, endospores, and biofilms [ 3 , 4 ]. Bacterial infections are posing an escalating threat to human health, with their treatment incurring substantial economic costs [ 5 ]. Currently, there is a paucity of research on drug molecules for the treatment of B. cereus . Among the few known synthetic antimicrobial compounds, the most potent ones exhibit a minimum inhibitory concentration of 0.97 µg/mL against B. cereus (as illustrated in Fig. 1 ) [ 6 – 9 ]. However, the inhibitory potency and target specificity of existing drugs warrant further enhancement. Consequently, there is an urgent need to increase research investment and continuously explore novel, highly effective, low-toxicity, and environmentally friendly inhibitors targeting B. cereus . 1,3,4-Oxadiazole derivatives have garnered significant interest due to their potential insecticidal, antimicrobial, anticancer, and anti-inflammatory activities, stemming from their unique structural properties [ 10 , 11 ]. Conversely, the incorporation of the trifluoromethyl (-CF 3 ) moiety into drug molecules can effectively modulate physicochemical properties and metabolic stability owing to its strong electron-withdrawing capability, lipophilicity, and robust C-F bond [ 12 – 14 ]. In this study, a "molecular hybridization" strategy was employed, combining antimicrobial norfloxacin derivatives and α -aminophosphonate derivatives, to design and synthesize a novel class of trifluoromethyl-1,3,4-oxadiazole amide antimicrobial compounds (as illustrated in Fig. 2 ) [ 15 , 16 ]. Molecular docking and molecular dynamics (MD) simulations enable the identification of precise geometric binding modes at ligand binding sites and the quantification of their binding affinities, thereby providing the necessary fundamental insights for rational drug design [ 17 ]. In this study, the antimicrobial potency was corroborated through theoretical models and some existing marketed antibiotics (such as Gentamicin and Ciprofloxacin). A synergistic approach involving molecular docking, MD simulations, chemical synthesis, and biological assays was employed to discover potent inhibitors targeting B. cereus . Results and discussion Chemistry synthesis For the synthesis of newly designed 1,3,4-oxadiazole-derived amide compounds, 3-(trifluoromethyl)benzaldehyde 1 was selected as the starting material, which can be directly procured from commercial suppliers. The specific synthetic route for these target compounds is illustrated in Scheme 1 . Initially, the semicarbazone intermediate 2 was readily obtained via a condensation reaction between benzaldehyde 1 and aminourea hydrochloride under basic catalysis, which upon I 2 -mediated intramolecular oxidative cyclization of semicarbazone 2 in alkaline medium afforded the desired 2-amino-substituted 1,3,4-oxadiazole skeleton intermediate 3 . Finally, the target 1,3,4-oxadiazole-derived amide compounds 1a-1n were synthesized by reacting intermediate 3 with various substituted aromatic carboxylic acids in the presence of EDCI as acid-amine coupling agent. Comprehensive structural characterization of all the target compounds was accomplished by 1 H-NMR, 13 C-NMR, HRMS, and FT-IR spectroscopic techniques. The obtained spectroscopic data were consistent with the assigned structures of the synthesized compounds, which were shown in Fig. S1-Fig. S14. Biological activity Antibacterial screening The potential antibacterial activities of compounds 1a-1b were initially evaluated against various bacterial strains (as shown in Table S1). Compounds 1a and 1b exhibited superior bactericidal effects against the 20 selected strains, reflecting their broad-spectrum antibacterial activity (as depicted in Fig. 3 ). Furthermore, compounds 1a and 1b under investigation demonstrated significant inhibitory activity against B. cereus FM314 ( 1a MIC = 2.5 µg/mL; 1b MIC = 0.15625 µg/mL), L. monocytogenes CMCC54002 ( 1a MIC = 5 µg/mL; 1b MIC = 0.625 µg/mL), S. aureus ATCC27660 ( 1a MIC = 2.5 µg/mL; 1b MIC = 0.15625 µg/mL), and C. albicans FM-2223 ( 1a MIC = 1.25 µg/mL; 1b MIC = 0.3125 µg/mL). The minimum inhibitory concentration (MIC) values reached the microgram level, and in some cases, even the nanogram level. Consequently, for subsequent compounds, the focus will be on evaluating and investigating the activity against these four strains. Table 1 Minimal inhibitory concentrations (MIC a , µg/mL) of 1a–1n against the tested B. cereus FM314、 L. monocytogenes CMCC54002、 S. aureus ATCC27660 and C. albicans FM-2223 strains. To comprehensively evaluate the antimicrobial activity of the target compounds, an in-depth investigation was conducted on four representative strains initially screened, with the aim of elucidating the inhibitory effects of these compounds against various pathogens. Ciprofloxacin and Gentamicin, two widely used antibiotics, were employed as positive controls for comparative analysis of the antimicrobial activity data. As illustrated in Fig. 4 , the vast majority of compounds exhibited remarkable antimicrobial potential at microgram concentration levels, with compounds 1c , 1d , 1i , 1j and 1n exhibiting minimum inhibitory concentrations (MICs) as low as 0.03907 µg/mL against the tested strains. Furthermore, the remaining compounds demonstrated potent inhibitory activity against B. cereus FM314, with antimicrobial efficacy surpassing or comparable to the positive control drugs (Gentamicin MIC = 0.625 µg/mL; Ciprofloxacin MIC = 2.5 µg/mL). Notably, the most potent drug molecules exhibited antimicrobial effects approximately 15-fold and 60-fold higher than Gentamicin and Ciprofloxacin, respectively, showcasing superior antimicrobial efficacy. Underpinned by the remarkable antimicrobial activity data, we hypothesized that the designed compounds might exhibit a characteristic inhibitory effect against the specific pathogen B. cereus . To gain in-depth understanding of the potential mechanism of action, we selected the most promising compounds 1c , 1d , 1i , 1j and 1n employed computational approaches, including molecular docking and molecular dynamics simulations, to systematically analyze their binding modes with the target bacterial strain. The objective was to elucidate the binding sites and interaction patterns of these lead compounds at the molecular level, thereby providing theoretical insights to guide subsequent structural optimization and activity enhancement endeavors. Structure-activity relationship (SAR) study Through structure-activity relationship (SAR) analysis, we discovered the pivotal influence of halogenated aromatic rings on antimicrobial activity. In comparison to compound 1a , the introduction of halogen substituents resulted in a remarkable enhancement of activity for the vast majority of compounds. Notably, the incorporation of halogen atoms at the 3rd and 4th positions of the phenyl ring further amplified the compounds' antimicrobial potential, with compound 1d (MIC = 0.0397 µg/mL) exhibiting approximately 4-fold higher activity than 1h (MIC = 0.15625 µg/mL). Another remarkable observation was that the introduction of a chlorine substituent at the 4th position of the phenyl ring substantially improved the antimicrobial activity of the compounds, such as compound 1e (MIC = 0.07813 µg/mL) > 1a (MIC = 2.5 µg/mL). Moreover, we evaluated the impact of steric hindrance on activity. The research findings indicated that as the volume of the substituent increased, the antimicrobial activity of the compounds concomitantly improved, as evident from the trend: 1g (MIC = 0.0397 µg/mL) > 1k (MIC = 0.7813 µg/mL) > 1b (MIC = 0.15625 µg/mL) > 1a (MIC = 2.5 µg/mL). This discovery provided a novel perspective for our subsequent drug design endeavors. Molecular docking results To elucidate the potential binding modes of the highly active compounds 1c , 1d , 1i , 1j and 1n with B. cereus , molecular docking studies were performed. The docking results of these compounds were compared against the target protein retrieved from the Protein Data Bank (PDB ID: 4EI9). Within the molecular docking simulations, the binding conformation exhibiting the highest docking score was prioritized for analysis. As depicted in Fig. 5 , within the active site cavity of the 4EI9 protein, the aforementioned small molecules exhibited potent binding affinities toward the target, engaging in various interactions, including hydrogen bonding and hydrophobic interactions. Notably, the majority of the targeted compounds formed robust hydrogen bond interactions with the amino acid residues Ser57 and Thr125, displaying binding energies that either surpassed or matched those of the reference drugs Gentamicin and Ciprofloxacin, with some compounds even demonstrating superior binding capabilities. Specifically, compounds 1c , 1d , 1i , 1j and 1n docked into the active site of 4EI9, yielding calculated binding energies of -10.92, -10.81, -11.68, -10.99 and − 11.76 kcal/mol, respectively (as shown in Fig. 5 ), indicative of their exceptional binding affinities toward the target protein. As presented in Table 2 , the binding energies and key interacting residues with the target protein for all targeted compounds. Molecular dynamics simulations and MMPBSA To gain a comprehensive understanding of the interaction dynamics between receptor proteins and small molecules, as well as assess the stability of the corresponding binding sites, we performed 100 ns molecular dynamics (MD) simulations on the compounds of interest. The root-mean-square deviation (RMSD) serves as a crucial parameter for evaluating whether complex biomolecular systems have attained equilibrium. As illustrated in Fig. 6 a, within the initial 20 ns timescale, the RMSD values of the individual systems exhibited pronounced fluctuations and perturbations, reflecting the structural adjustments and optimizations undertaken by the complexes. During this process, the proteins and ligands underwent mutual adaptations, seeking optimal binding conformations. As the simulations progressed, the RMSD values converged, beginning to stabilize within the range of 0.3–0.4 nm, indicating that the molecules had transitioned to a relatively equilibrated conformational state. Table 2 The docking affinity of the positive control drug and novel compounds. No. H-Bonds (Residues) Distance (Å) ΔG bind (kcal/mol) 1c Gly26,Ser57,Thr125 3.29,2.86,3.21 -10.92 1d Thr55,Gly124,Thr125 2.77,3.06,2.27 -10.81 1i Ser57,Thr125 2.94,3.15 -11.68 1j Ser57,Arg85 2.84,2.87,2.97,3.09 -10.99 1n Ser57,Glu58,Arg85,Thr125,Ser237 3.02,3.11,3.334,3.293.13 -11.76 Gentamicin Ser57,His233,Glu238 2.97,2.98,3.02 -10.87 Ciprofloxacin Ala154,Asp235,Arg333 3.03,2.85,2.83 -9.76 The root-mean-square fluctuation (RMSF) analysis serves as an effective quantitative tool for evaluating the conformational fluctuations and stability of individual residues within a protein. As illustrated in Fig. 6 b, while subtle differences exist among the various protein-ligand complex systems, the overall trends of the RMSF curves exhibit remarkable consistency. Notably, at certain specific amino acid residues, minor fluctuations in RMSF values were observed, potentially arising from local conformational adjustments induced by the distinct binding modes of the ligands. Significantly, the RMSF values for all residues were maintained below 0.8 nm, a relatively low level. The radius of gyration (Rg) is a widely utilized parameter for evaluating the compactness of protein structures, where smaller values indicate a more tightly packed and stable internal configuration. As depicted in Fig. 6 c, during the initial stages of the simulation, the Rg values of the individual systems exhibited fluctuations, reflecting the conformational adjustments undertaken by the proteins to accommodate ligand binding. Notably, as the simulation progressed, the Rg curves of all systems converged and ultimately stabilized within the range of approximately 3.20–3.25 nm. This numerical range suggests that the receptor protein maintained a relatively compact yet dynamic conformational state. Significantly, the differences in Rg values across the various systems gradually diminished, eventually nearly overlapping. Hydrogen bonding, a crucial non-covalent intermolecular interaction, plays a pivotal role in sustaining the binding between proteins and ligand molecules. As illustrated in Fig. 6 d, although subtle differences exist among the various complex systems, the number of hydrogen bonds formed in all systems remained within a relatively narrow fluctuation range throughout the simulation period, without exhibiting significant surges or declines. The binding free energy serves as a crucial metric for quantifying the interaction strength between small molecules and biological macromolecules. By precisely calculating the binding energies of different ligands and receptor proteins, we can gain profound insights into the inherent differences in their intermolecular interactions from an energetic perspective. As depicted in Table 3 , despite structural similarities, these six compounds exhibit notable variations in their binding energy values. Ciprofloxacin, employed as a positive control, exhibits the lowest binding energy (ΔG = -14.06 kcal/mol). In contrast, the 4EI9- 1n system displays a higher binding energy value (ΔG = -47.14 kcal/mol), implying a more stable and stronger interaction between the compound and the receptor protein, thereby explaining its superior antimicrobial activity. Notably, the binding energy of the 4EI9- 1j system is significantly lower than the other molecules, a disparity potentially stemming from conformational distortions experienced by this compound during the binding process, particularly conformational alterations in residues proximal to critical binding sites. Conclusion Herein, we report the design and synthesis of a novel series of molecules based on the 1,3,4-oxadiazole scaffold, which were subsequently evaluated for their antimicrobial activity against various target organisms. The majority of these compounds exhibited significant inhibitory effects against the Gram-positive bacterium B. cereus . To gain insights into the molecular basis of their bioactivity, we selected five most potent antimicrobial compounds and subjected them to molecular docking and MD simulations. Notably, the computational results were largely consistent with our experimental biological data, further validating the reliability of these in silico approaches. Importantly, through comprehensive structure-activity relationship (SAR) studies, we identified halogen substituents as a promising structural motif that can facilitate the synthesis of compounds with enhanced antimicrobial potency. These halogenated derivatives can be considered as promising lead candidates for further optimization and development of novel antimicrobial agents with improved efficacy and selectivity, thereby contributing to the ongoing efforts to combat the ever-increasing threat of bacterial resistance. Table 3 Combined free energy and different energy contributions. Complex ΔE ELE (kcal mol − 1 ) ΔE VDW (kcal mol − 1 ) ΔE VDW (kcal mol − 1 ) \(\varDelta {\varvec{G}}_{\varvec{S}\varvec{u}\varvec{r}\varvec{f}}\) (kcal mol − 1 ) ΔG bind (kcal mol − 1 ) Ciprofloxacin -0.76 ± 2.81 -22.20 ± 0.94 11.38 ± 0.45 -2.48 ± 0.42 -14.06 ± 3.00 Gentamicin -40.23 ± 4.61 -51.48 ± 1.02 59.20 ± 1.04 -6.55 ± 0.22 -39.06 ± 3.72 1c -13.41 ± 1.79 -41.87 ± 1.68 28.64 ± 1.86 -4.67 ± 0.17 -31.31 ± 1.93 1d -14.21 ± 1.85 -38.75 ± 2.10 26.38 ± 1.88 -4.35 ± 0.22 -30.92 ± 1.75 1i -32.85 ± 3.94 -52.04 ± 3.16 50.21 ± 2.44 -5.45 ± 0.19 -40.13 ± 3.40 1j -0.51 ± 0.72 -31.73 ± 4.28 16.12 ± 1.32 -3.66 ± 0.44 -19.79 ± 1.55 1n -9.59 ± 1.11 -35.95 ± 0.59 25.64 ± 0.89 -5.04 ± 0.55 -47.14 ± 1.16 Experimental Section All reagents were obtained from commercial sources (Adamas-Beta and Macklin) and purified by distillation or recrystallization to eliminate moisture effects. The compounds and reagents were purchased and ensured that all reagents were of analytical grade. Column chromatography was carried out using silica gel (200–300 mesh). Melting points were measured on a digital melting point apparatus (Zhuoguang GM30). Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker AVANCE III 500 MHz NMR spectrometer using DMSO- d 6 as the solvent and TMS as the internal standard. Mass spectra (HRMS) were conducted on Thermo Scientific Q Exactive Focus Mass Spectrometer. IR spectra were recorded on a Thermo FT-IR spectrometer (Nicolet IS20) and are reported in terms of frequency of absorption. Synthesis of 2-amino-substituted 1,3,4-oxadiazole intermediate 3 A solution of trifluoromethylbenzaldehyde (87 mg, 0.5 mmol) in methanol (0.5 mL) was added dropwise into the solution of semicarbazide hydrochloride (56 mg, 0.5 mmol) and sodium acetate (41 mg, 0.5 mmol) in H 2 O (1 mL) at 25 o C. After being stirred at the same temperature for 10 min, 1,4-dioxane (1.5 mL) was added in one portion to the resulting suspension. Then iodine (152 mg, 0.6 mmol) and K 2 CO 3 (207 mg, 1.5 mmol) were added and the resultant solution was heated to 80°C. After the reaction was completed (monitored by TLC, 12 h), a solution of 5% Na 2 S 2 O 3 (10 mL) is added to the reaction mixture at 25 o C and extracted with CH 2 Cl 2 /MeOH (10:1, 3×5 mL). The combined organic layer was dried over anhydrous MgSO 4 and concentrated under reduced pressure to give the crude product, which was purified by recrystallization (EA:PE = 1:2) to afford intermediate 3 as yellow solid (105 mg, 91%). m.p.: 229.1-229.8 o C (Lit.[ 18 ] m.p.: 229–230 o C); 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.09 (d, J = 8.0 Hz, 1H), 8.02 (s, 1H), 7.89 (d, J = 7.2 Hz, 1H), 7.79 (t, J = 8.0 Hz, 1H), 7.40 (br s, 2H); 13 C NMR (100 MHz, DMSO- d 6 ) δ 164.6, 156.6, 131.1, 130.3, 129.2, 127.2, 125.8, 124.1, 121.6; HRMS calculated for C 9 H 7 F 3 N 3 O [M + H] + 230.0536, found 230.0539. General procedure for the synthesis of trifluoromethyl-1,3,4-oxadiazole amide derivatives 1a-1n To a solution of intermediate 3 (1 mmol) in CH 2 Cl 2 (15 mL) was added EDCI (1 mmol), DMAP (1 mmol) and various substituted aromatic carboxylic acid (1 mmol). Then, the reaction mixture was heated to 45°C and stirred at reflux for 4–24 h (monitored by TLC). After cooling to room temperature, 5% HCl (12 mL) was added to the reaction mixture, the resulting suspension was stirred at room temperature for 10 min. The crystals that precipitated were collected by filtration and triturated with PE/AcOEt /MeOH (3:1:0.2, 5 mL) overnight, filtered and dried at 60 o C in vacuo for 2 h to give the target products 1a-1e , 1h , 1j , 1m and 1n . The other crude product ( 1f , 1g , 1i , 1k and 1l ) was purified by flash chromatography (AcOEt/PE = 1:2). In vitro antibacterial assay All strains were grown at 37°C for 12–16 h in LB until OD 600 = 0.6. Then bacteria were diluted 1:1000 into fresh medium containing TTC, the newly synthesized trifluoromethyl-1,3,4-oxadiazole amide compounds were dissolved in dimethyl sulfoxide (DMSO) to a concentration of 2 mg/mL, and the solutions were diluted with DMSO before antibacterial activity evaluation. For a comparison of antibacterial activity, the commercial gentamicin and ciprofloxacin were used as the reference drugs. Bacteria (198 µL) and compounds (2 µL) were added into the 96 well plates and mixed well by shaking. Then, the plates were incubated at 37°C for 12–16 h followed by observations of MIC values. For each compound, MIC determinations were performed independently thrice using duplicate samples each time. MIC was determined as the minimum concentration of compound at which visible bacterial growth is inhibited. Molecular dock methods In this study, we employed a semi-flexible docking approach to facilitate the formation of stable complexes. Initially, downloaded proteins containing co-crystallized ligands with structures similar to the synthesized compounds from the RCSB Protein Data Bank (PDB ID: 4EI9). The proteins were subsequently prepared using AutoDock Tools 1.5.2 software [ 19 ]. The preparation phase involved the addition of polar hydrogen atoms and the computation of partial charges. Thereafter, we performed a local charge distribution of the proteins and assigned AutoDock atom types. Subsequently, we established parameters in the Grid module by defining the docking region's box size as 126×126×126 to encompass the entire protein molecule, thereby aiming to locate the optimal binding site. The grid spacing was set to 0.375 Å, retaining grid information, while other parameters were kept as default [ 20 , 21 ]. The docking process was executed to generate the top ten optimal docking poses. Lastly, visualized the results using PyMol 2.0 software, enabling further analysis of the complex stability and intermolecular interactions. Molecular dynamics simulation and MMPBSA methods To gain in-depth insights into the intermolecular interactions and stability of the complexes, MD simulations were employed in this study. We simulated the protein-ligand complexes for 100 ns using the GROMACS 2021.3 software package [ 22 , 23 ]. The AMBER99SB-ILDN force field was selected to generate the protein topology files, while the Amber20 software and GAFF force field were utilized to generate the topology files for the small molecule ligands [ 24 ]. The systems were solvated with a truncated octahedral TIP3P water box extending 10 nm from the molecular surfaces, and Na/Cl ions were added to neutralize the net charge. Energy minimization was carried out using 2500 steps of the steepest descent method followed by 2500 steps of the conjugate gradient method. Subsequently, the systems were equilibrated under NVT conditions for 100 ps at 298.15 K, followed by 100 ps of NPT equilibration [ 25 , 26 ]. Finally, 100 ns of production MD simulations were performed under periodic boundary conditions using the NPT ensemble, with long-range electrostatic interactions calculated by the particle mesh Ewald (PME) method. The non-bonded cutoff distance was set to 1 nm, the collision frequency was 2 ps, the system pressure was maintained at 101.325 kPa, the integration time step was 2 fs, and coordinates were saved every 10 ps [ 27 ]. Furthermore, by extracting the stable molecular conformations from the last 20 ns of the equilibrated trajectories, employed the gmx_MMPBSA tool to calculate the binding free energies between the proteins and small molecules [ 28 , 29 ]. Through decomposes the binding free energy into individual energy components, such as van der Waals interactions, electrostatic interactions, and polar solvation energies, the relationship between small molecules and proteins was further analyzed from the perspective of energy. Declarations Author contributions Fei Xiong and Yanjun Zhang were in charge of writing this article and summarizing the data. Yanjun Zhang, Yiren Zhu and Yeji Li performed the synthetic experimental work, and characterization, and molecular dynamic simulation studies. Tianlu Mo and Jinlong Jiao carried out the in vitro antibacterial activity test. Fei Xiong conceived the project and provided the resources, supervision and funding assistance. All authors critically evaluated the manuscript prior to submission. Conflicts of interest There are no conflicts of interest to declare. Acknowledgements This work was supported by the financial support of the National Natural Science Foundation of China (NO. 81172918, 51707122), Shanghai Municipal Education Commission (NO. SLG14033) and the open project program of Hubei Key Laboratory of Drug Synthesis and Optimization, Jing Chu University of Technology (NO. OPP2014ZD01). We would like to thank Shiyanjia Lab (www.shiyanjia.com) for the support of NMR tests. References Cui P, Li X, Zhu M, Wang B et al (2017) Design, synthesis and antibacterial activities of thiouracil derivatives containing acyl thiourea as SecA inhibitors. Bioorg Med Chem Lett 27:2234–2237 Kanagasabapathy G, Britto S, Anbazhagan V (2023) Synthesis, characterization and molecular docking studies of highly functionalized and biologically active derivatives of 2-aminothiazole. J Mol Struct 1275:134593 Andersson DI, Hughes D, Kubicek-Sutherland JZ (2019) Mechanisms and consequences of bacterial resistance to antimicrobial peptides. 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J Chem Theory Comput 8:3314–3321 Scheme Scheme 1 is available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files Scheme1.png Scheme 1 Synthetic route of trifluoromethyl-1,3,4-oxadiazole amide derivatives. Reagent and conditions: (a) NaOAc, MeOH/H 2 O, semicarbazide hydrochloride, 25 o C, 1 h; (b) I 2 , K 2 CO 3 , 1,4-dioxane, 80 ℃, 12 h; (c) R-ArCO 2 H, EDCl, DMAP, dichloromethane, 45 ℃, 12 h. Cite Share Download PDF Status: Published Journal Publication published 20 Jun, 2024 Read the published version in Molecular Diversity → Version 1 posted Editorial decision: Revision requested 11 Apr, 2024 Reviews received at journal 06 Apr, 2024 Reviews received at journal 27 Mar, 2024 Reviewers agreed at journal 25 Mar, 2024 Reviewers agreed at journal 25 Mar, 2024 Reviewers invited by journal 25 Mar, 2024 Editor assigned by journal 22 Mar, 2024 Submission checks completed at journal 22 Mar, 2024 First submitted to journal 22 Mar, 2024 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. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4148372","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":283920881,"identity":"de7b83b5-b4b0-4998-8c71-ffdf9ba42cc2","order_by":0,"name":"Fei Xiong","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzklEQVRIiWNgGAWjYBAC9gbGBiBlA+WyEaGF5wBYSxoQMxOtBUwdJkWLRHLj44Jf5xPn958/wPCh7DAD/+wGAlp4DjYbz+y7nbjhRjID44xzhxkk7hzAr8WevbFNmrcHqEWCmYGZt+0wg4FEAgFbmBlBWs4BHXaYgfkvUVpAtvD8OJDYcCCZAaidGC0gv/A2JBsD/WJwsOdcOo/EDUJaJNIfPub5Yyc7v//gwwc/yqzl+GcQ0AIGjG0Q+gDIDCLUg8AfItWNglEwCkbByAQAUmVCb6VbriAAAAAASUVORK5CYII=","orcid":"","institution":"University of Shanghai for Science and Technology","correspondingAuthor":true,"prefix":"","firstName":"Fei","middleName":"","lastName":"Xiong","suffix":""},{"id":283920882,"identity":"77be95c2-9e1a-45b2-9732-412966b256ef","order_by":1,"name":"Yanjun Zhang","email":"","orcid":"","institution":"University of Shanghai for Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Yanjun","middleName":"","lastName":"Zhang","suffix":""},{"id":283920883,"identity":"49d39af8-3d26-42cc-b086-36ab437138d8","order_by":2,"name":"Jinlong Jiao","email":"","orcid":"","institution":"University of Shanghai for Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Jinlong","middleName":"","lastName":"Jiao","suffix":""},{"id":283920884,"identity":"d6ee9fb3-9510-4cb3-9233-169a56bafa0e","order_by":3,"name":"Yiren Zhu","email":"","orcid":"","institution":"University of Shanghai for Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Yiren","middleName":"","lastName":"Zhu","suffix":""},{"id":283920885,"identity":"5c0bec69-f823-444c-8e36-96ac630824c7","order_by":4,"name":"Tianlu Mo","email":"","orcid":"","institution":"University of Shanghai for Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Tianlu","middleName":"","lastName":"Mo","suffix":""},{"id":283920886,"identity":"03ec0e23-5426-4954-95a1-80e4f6715f5c","order_by":5,"name":"Yeji Li","email":"","orcid":"","institution":"University of Shanghai for Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Yeji","middleName":"","lastName":"Li","suffix":""}],"badges":[],"createdAt":"2024-03-22 08:54:48","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4148372/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4148372/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11030-024-10893-x","type":"published","date":"2024-06-20T16:10:42+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":53601480,"identity":"4082cd73-4ce1-4f6c-8a09-6315bf3fc9eb","added_by":"auto","created_at":"2024-03-28 02:02:58","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":43494,"visible":true,"origin":"","legend":"\u003cp\u003eStructures of \u003cem\u003eBacillus cereus\u003c/em\u003e inhibitors with biological activities reported in the literature.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4148372/v1/e2913fd3f1aa660758910057.png"},{"id":53601617,"identity":"91db20a1-7627-4ec1-b543-cf19fce7e416","added_by":"auto","created_at":"2024-03-28 02:10:59","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":24025,"visible":true,"origin":"","legend":"\u003cp\u003eDesign strategy of the target trifluoromethyl-1,3,4-oxadiazole amide derivatives.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4148372/v1/f4e595e0c3353ca4f88f9cf0.png"},{"id":53601479,"identity":"07c04239-3b61-4d3d-9d3b-f5d6a989d113","added_by":"auto","created_at":"2024-03-28 02:02:58","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":102959,"visible":true,"origin":"","legend":"\u003cp\u003eMIC values of compounds \u003cstrong\u003e1a\u003c/strong\u003e and \u003cstrong\u003e1b\u003c/strong\u003e against antibacterial activity. The specific values were shown in Table S1.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4148372/v1/871bc5be4c3e323df60df75f.png"},{"id":53601481,"identity":"3fc82bbd-653a-459c-9d74-d5f37621fa1a","added_by":"auto","created_at":"2024-03-28 02:02:58","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":125919,"visible":true,"origin":"","legend":"\u003cp\u003eAntibacterial activity of tested compounds \u003cem\u003eB. cereus \u003c/em\u003eFM314、\u003cem\u003eL. monocytogenes \u003c/em\u003eCMCC54002、\u003cem\u003eS. aureus \u003c/em\u003eATCC27660 and \u003cem\u003eC. albicans \u003c/em\u003eFM-2223 strains expressed as MICs (µg/mL).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4148372/v1/19d3a20a3e799426847a01eb.png"},{"id":53601482,"identity":"0f98ef72-0adb-4fa8-8077-1bdf8d390cd2","added_by":"auto","created_at":"2024-03-28 02:02:58","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":274222,"visible":true,"origin":"","legend":"\u003cp\u003eThe docking result of (1a, 1b) compound \u003cstrong\u003e1c\u003c/strong\u003e, (2a,2b) compound \u003cstrong\u003e1d\u003c/strong\u003e, (3a,3b) compound \u003cstrong\u003e1i\u003c/strong\u003e, (4a,4b) compound \u003cstrong\u003e1j\u003c/strong\u003e, (5a,5b) compound \u003cstrong\u003e1n\u003c/strong\u003e, (6a,6b) compound Gentamicin and (7a,7b) compound Ciprofloxacin into the binding site of receptor protein (hydrogen bonds are shown as yellow dashed lines).\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-4148372/v1/077cd5a56f6c4b9aa763c9b2.png"},{"id":53601484,"identity":"3dfb1f26-ada3-4b80-8843-9ce7d19f84d0","added_by":"auto","created_at":"2024-03-28 02:02:59","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":239803,"visible":true,"origin":"","legend":"\u003cp\u003eThe results of 100 ns MD simulations of complexes 4EI9-Ciprofloxacin (black), 4EI9-Gentamicin (red), 4EI9-\u003cstrong\u003e1c\u003c/strong\u003e (blue), 4EI9-\u003cstrong\u003e1d\u003c/strong\u003e (green), 4EI9-\u003cstrong\u003e1i\u003c/strong\u003e(purple), 4EI9-\u003cstrong\u003e1j \u003c/strong\u003e(yellow), and 4EI9-\u003cstrong\u003e1n\u003c/strong\u003e (cyan). (a) The RMSD values of complex systems. (b) The RMSF values of residues in protein-ligand complexes. (c)The Rg values of complex systems. (d) The hydrogen bond number of complex systems.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-4148372/v1/c676bf13d4436924f79badbe.png"},{"id":58824030,"identity":"68fa2b3e-5201-4e19-ace7-32afe0e6e45a","added_by":"auto","created_at":"2024-06-21 17:13:51","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1454418,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4148372/v1/3def2af7-d307-458a-bc21-f7e8fffb7ea5.pdf"},{"id":53601483,"identity":"e5678d3b-0a0e-4d2c-b6c5-2dd14108089e","added_by":"auto","created_at":"2024-03-28 02:02:59","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":19517,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eScheme 1\u003c/strong\u003e Synthetic route of trifluoromethyl-1,3,4-oxadiazole amide derivatives. \u003cstrong\u003eReagent and conditions\u003c/strong\u003e: (a) NaOAc, MeOH/H\u003csub\u003e2\u003c/sub\u003eO, semicarbazide hydrochloride, 25 \u003csup\u003eo\u003c/sup\u003eC, 1 h; (b) I\u003csub\u003e2\u003c/sub\u003e, K\u003csub\u003e2\u003c/sub\u003eCO\u003csub\u003e3\u003c/sub\u003e, 1,4-dioxane, 80 ℃, 12 h; (c) R-ArCO\u003csub\u003e2\u003c/sub\u003eH, EDCl, DMAP, dichloromethane, 45 ℃, 12 h.\u003c/p\u003e","description":"","filename":"Scheme1.png","url":"https://assets-eu.researchsquare.com/files/rs-4148372/v1/75f29c3e8f2dbe00db3ae3f3.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Towards new bioactive fluorine-containing 1,3,4-oxadiazole-amide derivatives: Synthesis, antibacterial activity, molecular docking and molecular dynamics simulation study","fulltext":[{"header":"Introduction","content":"\u003cp\u003eBacterial diseases stemming from escalating antimicrobial resistance have emerged as a paramount concern in the public health domain in recent years [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. The ubiquitous foodborne pathogen-\u003cem\u003eBacillus cereus\u003c/em\u003e, which contaminates a range of edible products, including flour-based items and dairy products, poses a significant health threat to both adults and children [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Research has elucidated that this bacterial species is ubiquitous in diverse environments, such as soil, water, air, and food matrices, existing in various forms, including vegetative cells, endospores, and biofilms [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eBacterial infections are posing an escalating threat to human health, with their treatment incurring substantial economic costs [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Currently, there is a paucity of research on drug molecules for the treatment of \u003cem\u003eB. cereus\u003c/em\u003e. Among the few known synthetic antimicrobial compounds, the most potent ones exhibit a minimum inhibitory concentration of 0.97 \u0026micro;g/mL against \u003cem\u003eB. cereus\u003c/em\u003e (as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) [\u003cspan additionalcitationids=\"CR7 CR8\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. However, the inhibitory potency and target specificity of existing drugs warrant further enhancement. Consequently, there is an urgent need to increase research investment and continuously explore novel, highly effective, low-toxicity, and environmentally friendly inhibitors targeting \u003cem\u003eB. cereus\u003c/em\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e1,3,4-Oxadiazole derivatives have garnered significant interest due to their potential insecticidal, antimicrobial, anticancer, and anti-inflammatory activities, stemming from their unique structural properties [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Conversely, the incorporation of the trifluoromethyl (-CF\u003csub\u003e3\u003c/sub\u003e) moiety into drug molecules can effectively modulate physicochemical properties and metabolic stability owing to its strong electron-withdrawing capability, lipophilicity, and robust C-F bond [\u003cspan additionalcitationids=\"CR13\" citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. In this study, a \"molecular hybridization\" strategy was employed, combining antimicrobial norfloxacin derivatives and \u003cem\u003eα\u003c/em\u003e-aminophosphonate derivatives, to design and synthesize a novel class of trifluoromethyl-1,3,4-oxadiazole amide antimicrobial compounds (as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMolecular docking and molecular dynamics (MD) simulations enable the identification of precise geometric binding modes at ligand binding sites and the quantification of their binding affinities, thereby providing the necessary fundamental insights for rational drug design [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. In this study, the antimicrobial potency was corroborated through theoretical models and some existing marketed antibiotics (such as Gentamicin and Ciprofloxacin). A synergistic approach involving molecular docking, MD simulations, chemical synthesis, and biological assays was employed to discover potent inhibitors targeting \u003cem\u003eB. cereus\u003c/em\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Results and discussion","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003eChemistry synthesis\u003c/h2\u003e\n \u003cp\u003eFor the synthesis of newly designed 1,3,4-oxadiazole-derived amide compounds, 3-(trifluoromethyl)benzaldehyde \u003cstrong\u003e1\u003c/strong\u003e was selected as the starting material, which can be directly procured from commercial suppliers. The specific synthetic route for these target compounds is illustrated in Scheme \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. Initially, the semicarbazone intermediate \u003cstrong\u003e2\u003c/strong\u003e was readily obtained via a condensation reaction between benzaldehyde \u003cstrong\u003e1\u003c/strong\u003e and aminourea hydrochloride under basic catalysis, which upon I\u003csub\u003e2\u003c/sub\u003e-mediated intramolecular oxidative cyclization of semicarbazone \u003cstrong\u003e2\u003c/strong\u003e in alkaline medium afforded the desired 2-amino-substituted 1,3,4-oxadiazole skeleton intermediate \u003cstrong\u003e3\u003c/strong\u003e. Finally, the target 1,3,4-oxadiazole-derived amide compounds \u003cstrong\u003e1a-1n\u003c/strong\u003e were synthesized by reacting intermediate \u003cstrong\u003e3\u003c/strong\u003e with various substituted aromatic carboxylic acids in the presence of EDCI as acid-amine coupling agent. Comprehensive structural characterization of all the target compounds was accomplished by \u003csup\u003e1\u003c/sup\u003eH-NMR, \u003csup\u003e13\u003c/sup\u003eC-NMR, HRMS, and FT-IR spectroscopic techniques. The obtained spectroscopic data were consistent with the assigned structures of the synthesized compounds, which were shown in Fig. S1-Fig. S14.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n \u003ch2\u003eBiological activity\u003c/h2\u003e\n \u003cdiv id=\"Sec5\" class=\"Section3\"\u003e\n \u003ch2\u003eAntibacterial screening\u003c/h2\u003e\n \u003cp\u003eThe potential antibacterial activities of compounds \u003cstrong\u003e1a-1b\u003c/strong\u003e were initially evaluated against various bacterial strains (as shown in Table S1). Compounds \u003cstrong\u003e1a\u003c/strong\u003e and \u003cstrong\u003e1b\u003c/strong\u003e exhibited superior bactericidal effects against the 20 selected strains, reflecting their broad-spectrum antibacterial activity (as depicted in Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). Furthermore, compounds \u003cstrong\u003e1a\u003c/strong\u003e and \u003cstrong\u003e1b\u003c/strong\u003e under investigation demonstrated significant inhibitory activity against \u003cem\u003eB. cereus\u003c/em\u003e FM314 (\u003cstrong\u003e1a\u003c/strong\u003e MIC\u0026thinsp;=\u0026thinsp;2.5 \u0026micro;g/mL; \u003cstrong\u003e1b\u003c/strong\u003e MIC\u0026thinsp;=\u0026thinsp;0.15625 \u0026micro;g/mL), \u003cem\u003eL. monocytogenes\u003c/em\u003e CMCC54002 (\u003cstrong\u003e1a\u003c/strong\u003e MIC\u0026thinsp;=\u0026thinsp;5 \u0026micro;g/mL; \u003cstrong\u003e1b\u003c/strong\u003e MIC\u0026thinsp;=\u0026thinsp;0.625 \u0026micro;g/mL), \u003cem\u003eS. aureus\u003c/em\u003e ATCC27660 (\u003cstrong\u003e1a\u003c/strong\u003e MIC\u0026thinsp;=\u0026thinsp;2.5 \u0026micro;g/mL; \u003cstrong\u003e1b\u003c/strong\u003e MIC\u0026thinsp;=\u0026thinsp;0.15625 \u0026micro;g/mL), and \u003cem\u003eC. albicans\u003c/em\u003e FM-2223 (\u003cstrong\u003e1a\u003c/strong\u003e MIC\u0026thinsp;=\u0026thinsp;1.25 \u0026micro;g/mL; \u003cstrong\u003e1b\u003c/strong\u003e MIC\u0026thinsp;=\u0026thinsp;0.3125 \u0026micro;g/mL). The minimum inhibitory concentration (MIC) values reached the microgram level, and in some cases, even the nanogram level. Consequently, for subsequent compounds, the focus will be on evaluating and investigating the activity against these four strains.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e Minimal inhibitory concentrations (MIC\u003csup\u003ea\u003c/sup\u003e, \u0026micro;g/mL) of \u003cstrong\u003e1a\u0026ndash;1n\u003c/strong\u003e against the tested \u003cem\u003eB. cereus\u0026nbsp;\u003c/em\u003eFM314、\u003cem\u003eL. monocytogenes\u0026nbsp;\u003c/em\u003eCMCC54002、\u003cem\u003eS. aureus\u0026nbsp;\u003c/em\u003eATCC27660 and \u003cem\u003eC. albicans\u003c/em\u003e FM-2223 strains.\u003c/p\u003e\n \u003cp\u003e\u003cimg 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\"\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003eTo comprehensively evaluate the antimicrobial activity of the target compounds, an in-depth investigation was conducted on four representative strains initially screened, with the aim of elucidating the inhibitory effects of these compounds against various pathogens. Ciprofloxacin and Gentamicin, two widely used antibiotics, were employed as positive controls for comparative analysis of the antimicrobial activity data. As illustrated in Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e, the vast majority of compounds exhibited remarkable antimicrobial potential at microgram concentration levels, with compounds \u003cstrong\u003e1c\u003c/strong\u003e, \u003cstrong\u003e1d\u003c/strong\u003e, \u003cstrong\u003e1i\u003c/strong\u003e, \u003cstrong\u003e1j\u003c/strong\u003e and \u003cstrong\u003e1n\u003c/strong\u003e exhibiting minimum inhibitory concentrations (MICs) as low as 0.03907 \u0026micro;g/mL against the tested strains. Furthermore, the remaining compounds demonstrated potent inhibitory activity against \u003cem\u003eB. cereus\u003c/em\u003e FM314, with antimicrobial efficacy surpassing or comparable to the positive control drugs (Gentamicin MIC\u0026thinsp;=\u0026thinsp;0.625 \u0026micro;g/mL; Ciprofloxacin MIC\u0026thinsp;=\u0026thinsp;2.5 \u0026micro;g/mL). Notably, the most potent drug molecules exhibited antimicrobial effects approximately 15-fold and 60-fold higher than Gentamicin and Ciprofloxacin, respectively, showcasing superior antimicrobial efficacy.\u003c/div\u003e\n \u003c/div\u003e\n \u003cp\u003eUnderpinned by the remarkable antimicrobial activity data, we hypothesized that the designed compounds might exhibit a characteristic inhibitory effect against the specific pathogen \u003cem\u003eB. cereus\u003c/em\u003e. To gain in-depth understanding of the potential mechanism of action, we selected the most promising compounds \u003cstrong\u003e1c\u003c/strong\u003e, \u003cstrong\u003e1d\u003c/strong\u003e, \u003cstrong\u003e1i\u003c/strong\u003e, \u003cstrong\u003e1j\u003c/strong\u003e and \u003cstrong\u003e1n\u003c/strong\u003e employed computational approaches, including molecular docking and molecular dynamics simulations, to systematically analyze their binding modes with the target bacterial strain. The objective was to elucidate the binding sites and interaction patterns of these lead compounds at the molecular level, thereby providing theoretical insights to guide subsequent structural optimization and activity enhancement endeavors.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n \u003ch2\u003eStructure-activity relationship (SAR) study\u003c/h2\u003e\n \u003cp\u003eThrough structure-activity relationship (SAR) analysis, we discovered the pivotal influence of halogenated aromatic rings on antimicrobial activity. In comparison to compound \u003cstrong\u003e1a\u003c/strong\u003e, the introduction of halogen substituents resulted in a remarkable enhancement of activity for the vast majority of compounds. Notably, the incorporation of halogen atoms at the 3rd and 4th positions of the phenyl ring further amplified the compounds\u0026apos; antimicrobial potential, with compound \u003cstrong\u003e1d\u003c/strong\u003e (MIC\u0026thinsp;=\u0026thinsp;0.0397 \u0026micro;g/mL) exhibiting approximately 4-fold higher activity than \u003cstrong\u003e1h\u003c/strong\u003e (MIC\u0026thinsp;=\u0026thinsp;0.15625 \u0026micro;g/mL). Another remarkable observation was that the introduction of a chlorine substituent at the 4th position of the phenyl ring substantially improved the antimicrobial activity of the compounds, such as compound \u003cstrong\u003e1e\u003c/strong\u003e (MIC\u0026thinsp;=\u0026thinsp;0.07813 \u0026micro;g/mL)\u0026thinsp;\u0026gt;\u0026thinsp;\u003cstrong\u003e1a\u003c/strong\u003e (MIC\u0026thinsp;=\u0026thinsp;2.5 \u0026micro;g/mL). Moreover, we evaluated the impact of steric hindrance on activity. The research findings indicated that as the volume of the substituent increased, the antimicrobial activity of the compounds concomitantly improved, as evident from the trend: \u003cstrong\u003e1g\u003c/strong\u003e (MIC\u0026thinsp;=\u0026thinsp;0.0397 \u0026micro;g/mL)\u0026thinsp;\u0026gt;\u0026thinsp;\u003cstrong\u003e1k\u003c/strong\u003e (MIC\u0026thinsp;=\u0026thinsp;0.7813 \u0026micro;g/mL)\u0026thinsp;\u0026gt;\u0026thinsp;\u003cstrong\u003e1b\u003c/strong\u003e (MIC\u0026thinsp;=\u0026thinsp;0.15625 \u0026micro;g/mL)\u0026thinsp;\u0026gt;\u0026thinsp;\u003cstrong\u003e1a\u003c/strong\u003e (MIC\u0026thinsp;=\u0026thinsp;2.5 \u0026micro;g/mL). This discovery provided a novel perspective for our subsequent drug design endeavors.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n \u003ch2\u003eMolecular docking results\u003c/h2\u003e\n \u003cp\u003eTo elucidate the potential binding modes of the highly active compounds \u003cstrong\u003e1c\u003c/strong\u003e, \u003cstrong\u003e1d\u003c/strong\u003e, \u003cstrong\u003e1i\u003c/strong\u003e, \u003cstrong\u003e1j\u003c/strong\u003e and \u003cstrong\u003e1n\u003c/strong\u003e with \u003cem\u003eB. cereus\u003c/em\u003e, molecular docking studies were performed. The docking results of these compounds were compared against the target protein retrieved from the Protein Data Bank (PDB ID: 4EI9). Within the molecular docking simulations, the binding conformation exhibiting the highest docking score was prioritized for analysis.\u003c/p\u003e\n \u003cp\u003eAs depicted in Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e, within the active site cavity of the 4EI9 protein, the aforementioned small molecules exhibited potent binding affinities toward the target, engaging in various interactions, including hydrogen bonding and hydrophobic interactions. Notably, the majority of the targeted compounds formed robust hydrogen bond interactions with the amino acid residues Ser57 and Thr125, displaying binding energies that either surpassed or matched those of the reference drugs Gentamicin and Ciprofloxacin, with some compounds even demonstrating superior binding capabilities. Specifically, compounds \u003cstrong\u003e1c\u003c/strong\u003e, \u003cstrong\u003e1d\u003c/strong\u003e, \u003cstrong\u003e1i\u003c/strong\u003e, \u003cstrong\u003e1j\u003c/strong\u003e and \u003cstrong\u003e1n\u003c/strong\u003e docked into the active site of 4EI9, yielding calculated binding energies of -10.92, -10.81, -11.68, -10.99 and \u0026minus;\u0026thinsp;11.76 kcal/mol, respectively (as shown in Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e), indicative of their exceptional binding affinities toward the target protein. As presented in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, the binding energies and key interacting residues with the target protein for all targeted compounds.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n \u003ch2\u003eMolecular dynamics simulations and MMPBSA\u003c/h2\u003e\n \u003cp\u003eTo gain a comprehensive understanding of the interaction dynamics between receptor proteins and small molecules, as well as assess the stability of the corresponding binding sites, we performed 100 ns molecular dynamics (MD) simulations on the compounds of interest. The root-mean-square deviation (RMSD) serves as a crucial parameter for evaluating whether complex biomolecular systems have attained equilibrium. As illustrated in Fig. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003ea, within the initial 20 ns timescale, the RMSD values of the individual systems exhibited pronounced fluctuations and perturbations, reflecting the structural adjustments and optimizations undertaken by the complexes. During this process, the proteins and ligands underwent mutual adaptations, seeking optimal binding conformations. As the simulations progressed, the RMSD values converged, beginning to stabilize within the range of 0.3\u0026ndash;0.4 nm, indicating that the molecules had transitioned to a relatively equilibrated conformational state.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eThe docking affinity of the positive control drug and novel compounds.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"4\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eNo.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eH-Bonds (Residues)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDistance (\u0026Aring;)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u0026Delta;G\u003csub\u003ebind\u003c/sub\u003e (kcal/mol)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e1c\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGly26,Ser57,Thr125\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.29,2.86,3.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e-10.92\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e1d\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eThr55,Gly124,Thr125\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.77,3.06,2.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e-10.81\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e1i\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSer57,Thr125\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.94,3.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e-11.68\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e1j\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSer57,Arg85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.84,2.87,2.97,3.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e-10.99\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e1n\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSer57,Glu58,Arg85,Thr125,Ser237\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.02,3.11,3.334,3.293.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e-11.76\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eGentamicin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSer57,His233,Glu238\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.97,2.98,3.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e-10.87\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eCiprofloxacin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAla154,Asp235,Arg333\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.03,2.85,2.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e-9.76\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eThe root-mean-square fluctuation (RMSF) analysis serves as an effective quantitative tool for evaluating the conformational fluctuations and stability of individual residues within a protein. As illustrated in Fig. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003eb, while subtle differences exist among the various protein-ligand complex systems, the overall trends of the RMSF curves exhibit remarkable consistency. Notably, at certain specific amino acid residues, minor fluctuations in RMSF values were observed, potentially arising from local conformational adjustments induced by the distinct binding modes of the ligands. Significantly, the RMSF values for all residues were maintained below 0.8 nm, a relatively low level.\u003c/p\u003e\n \u003cp\u003eThe radius of gyration (Rg) is a widely utilized parameter for evaluating the compactness of protein structures, where smaller values indicate a more tightly packed and stable internal configuration. As depicted in Fig. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003ec, during the initial stages of the simulation, the Rg values of the individual systems exhibited fluctuations, reflecting the conformational adjustments undertaken by the proteins to accommodate ligand binding. Notably, as the simulation progressed, the Rg curves of all systems converged and ultimately stabilized within the range of approximately 3.20\u0026ndash;3.25 nm. This numerical range suggests that the receptor protein maintained a relatively compact yet dynamic conformational state. Significantly, the differences in Rg values across the various systems gradually diminished, eventually nearly overlapping.\u003c/p\u003e\n \u003cp\u003eHydrogen bonding, a crucial non-covalent intermolecular interaction, plays a pivotal role in sustaining the binding between proteins and ligand molecules. As illustrated in Fig. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003ed, although subtle differences exist among the various complex systems, the number of hydrogen bonds formed in all systems remained within a relatively narrow fluctuation range throughout the simulation period, without exhibiting significant surges or declines.\u003c/p\u003e\n \u003cp\u003eThe binding free energy serves as a crucial metric for quantifying the interaction strength between small molecules and biological macromolecules. By precisely calculating the binding energies of different ligands and receptor proteins, we can gain profound insights into the inherent differences in their intermolecular interactions from an energetic perspective. As depicted in Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e, despite structural similarities, these six compounds exhibit notable variations in their binding energy values. Ciprofloxacin, employed as a positive control, exhibits the lowest binding energy (\u0026Delta;G = -14.06 kcal/mol). In contrast, the 4EI9-\u003cstrong\u003e1n\u003c/strong\u003e system displays a higher binding energy value (\u0026Delta;G = -47.14 kcal/mol), implying a more stable and stronger interaction between the compound and the receptor protein, thereby explaining its superior antimicrobial activity. Notably, the binding energy of the 4EI9-\u003cstrong\u003e1j\u003c/strong\u003e system is significantly lower than the other molecules, a disparity potentially stemming from conformational distortions experienced by this compound during the binding process, particularly conformational alterations in residues proximal to critical binding sites.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eHerein, we report the design and synthesis of a novel series of molecules based on the 1,3,4-oxadiazole scaffold, which were subsequently evaluated for their antimicrobial activity against various target organisms. The majority of these compounds exhibited significant inhibitory effects against the Gram-positive bacterium \u003cem\u003eB. cereus\u003c/em\u003e. To gain insights into the molecular basis of their bioactivity, we selected five most potent antimicrobial compounds and subjected them to molecular docking and MD simulations. Notably, the computational results were largely consistent with our experimental biological data, further validating the reliability of these in silico approaches. Importantly, through comprehensive structure-activity relationship (SAR) studies, we identified halogen substituents as a promising structural motif that can facilitate the synthesis of compounds with enhanced antimicrobial potency. These halogenated derivatives can be considered as promising lead candidates for further optimization and development of novel antimicrobial agents with improved efficacy and selectivity, thereby contributing to the ongoing efforts to combat the ever-increasing threat of bacterial resistance.\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\u003eCombined free energy and different energy contributions.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eComplex\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eΔE\u003csub\u003eELE\u003c/sub\u003e (kcal mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eΔE\u003csub\u003eVDW\u003c/sub\u003e (kcal mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eΔE\u003csub\u003eVDW\u003c/sub\u003e (kcal mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\varDelta {\\varvec{G}}_{\\varvec{S}\\varvec{u}\\varvec{r}\\varvec{f}}\\)\u003c/span\u003e\u003c/span\u003e(kcal mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eΔG\u003csub\u003ebind\u003c/sub\u003e(kcal mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\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\u003eCiprofloxacin\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.76\u0026thinsp;\u0026plusmn;\u0026thinsp;2.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e-22.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e11.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e-2.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e-14.06\u0026thinsp;\u0026plusmn;\u0026thinsp;3.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGentamicin\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-40.23\u0026thinsp;\u0026plusmn;\u0026thinsp;4.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e-51.48\u0026thinsp;\u0026plusmn;\u0026thinsp;1.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e59.20\u0026thinsp;\u0026plusmn;\u0026thinsp;1.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e-6.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e-39.06\u0026thinsp;\u0026plusmn;\u0026thinsp;3.72\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e1c\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-13.41\u0026thinsp;\u0026plusmn;\u0026thinsp;1.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e-41.87\u0026thinsp;\u0026plusmn;\u0026thinsp;1.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e28.64\u0026thinsp;\u0026plusmn;\u0026thinsp;1.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e-4.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e-31.31\u0026thinsp;\u0026plusmn;\u0026thinsp;1.93\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e1d\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-14.21\u0026thinsp;\u0026plusmn;\u0026thinsp;1.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e-38.75\u0026thinsp;\u0026plusmn;\u0026thinsp;2.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e26.38\u0026thinsp;\u0026plusmn;\u0026thinsp;1.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e-4.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e-30.92\u0026thinsp;\u0026plusmn;\u0026thinsp;1.75\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e1i\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-32.85\u0026thinsp;\u0026plusmn;\u0026thinsp;3.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e-52.04\u0026thinsp;\u0026plusmn;\u0026thinsp;3.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e50.21\u0026thinsp;\u0026plusmn;\u0026thinsp;2.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e-5.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e-40.13\u0026thinsp;\u0026plusmn;\u0026thinsp;3.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e1j\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-0.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e-31.73\u0026thinsp;\u0026plusmn;\u0026thinsp;4.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e16.12\u0026thinsp;\u0026plusmn;\u0026thinsp;1.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e-3.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e-19.79\u0026thinsp;\u0026plusmn;\u0026thinsp;1.55\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e1n\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e-9.59\u0026thinsp;\u0026plusmn;\u0026thinsp;1.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e-35.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e25.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e-5.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e-47.14\u0026thinsp;\u0026plusmn;\u0026thinsp;1.16\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e"},{"header":"Experimental Section","content":"\u003cp\u003eAll reagents were obtained from commercial sources (Adamas-Beta and Macklin) and purified by distillation or recrystallization to eliminate moisture effects. The compounds and reagents were purchased and ensured that all reagents were of analytical grade. Column chromatography was carried out using silica gel (200\u0026ndash;300 mesh). Melting points were measured on a digital melting point apparatus (Zhuoguang GM30). Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker AVANCE III 500 MHz NMR spectrometer using DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e as the solvent and TMS as the internal standard. Mass spectra (HRMS) were conducted on Thermo Scientific Q Exactive Focus Mass Spectrometer. IR spectra were recorded on a Thermo FT-IR spectrometer (Nicolet IS20) and are reported in terms of frequency of absorption.\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003eSynthesis of 2-amino-substituted 1,3,4-oxadiazole intermediate 3\u003c/b\u003e\u003c/h2\u003e \u003cp\u003eA solution of trifluoromethylbenzaldehyde (87 mg, 0.5 mmol) in methanol (0.5 mL) was added dropwise into the solution of semicarbazide hydrochloride (56 mg, 0.5 mmol) and sodium acetate (41 mg, 0.5 mmol) in H\u003csub\u003e2\u003c/sub\u003eO (1 mL) at 25 \u003csup\u003eo\u003c/sup\u003eC. After being stirred at the same temperature for 10 min, 1,4-dioxane (1.5 mL) was added in one portion to the resulting suspension. Then iodine (152 mg, 0.6 mmol) and K\u003csub\u003e2\u003c/sub\u003eCO\u003csub\u003e3\u003c/sub\u003e (207 mg, 1.5 mmol) were added and the resultant solution was heated to 80\u0026deg;C. After the reaction was completed (monitored by TLC, 12 h), a solution of 5% Na\u003csub\u003e2\u003c/sub\u003eS\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e (10 mL) is added to the reaction mixture at 25 \u003csup\u003eo\u003c/sup\u003eC and extracted with CH\u003csub\u003e2\u003c/sub\u003eCl\u003csub\u003e2\u003c/sub\u003e/MeOH (10:1, 3\u0026times;5 mL). The combined organic layer was dried over anhydrous MgSO\u003csub\u003e4\u003c/sub\u003e and concentrated under reduced pressure to give the crude product, which was purified by recrystallization (EA:PE\u0026thinsp;=\u0026thinsp;1:2) to afford intermediate \u003cb\u003e3\u003c/b\u003e as yellow solid (105 mg, 91%). m.p.: 229.1-229.8 \u003csup\u003eo\u003c/sup\u003eC (Lit.[\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e] m.p.: 229\u0026ndash;230 \u003csup\u003eo\u003c/sup\u003eC); \u003csup\u003e1\u003c/sup\u003eH NMR (400 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e) \u003cem\u003eδ\u003c/em\u003e 8.09 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.0 Hz, 1H), 8.02 (s, 1H), 7.89 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.2 Hz, 1H), 7.79 (t, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.0 Hz, 1H), 7.40 (br s, 2H); \u003csup\u003e13\u003c/sup\u003eC NMR (100 MHz, DMSO-\u003cem\u003ed\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e) \u003cem\u003eδ\u003c/em\u003e 164.6, 156.6, 131.1, 130.3, 129.2, 127.2, 125.8, 124.1, 121.6; HRMS calculated for C\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e7\u003c/sub\u003eF\u003csub\u003e3\u003c/sub\u003eN\u003csub\u003e3\u003c/sub\u003eO [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e 230.0536, found 230.0539.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003eGeneral procedure for the synthesis of trifluoromethyl-1,3,4-oxadiazole amide derivatives 1a-1n\u003c/b\u003e\u003c/h2\u003e \u003cp\u003eTo a solution of intermediate \u003cb\u003e3\u003c/b\u003e (1 mmol) in CH\u003csub\u003e2\u003c/sub\u003eCl\u003csub\u003e2\u003c/sub\u003e (15 mL) was added EDCI (1 mmol), DMAP (1 mmol) and various substituted aromatic carboxylic acid (1 mmol). Then, the reaction mixture was heated to 45\u0026deg;C and stirred at reflux for 4\u0026ndash;24 h (monitored by TLC). After cooling to room temperature, 5% HCl (12 mL) was added to the reaction mixture, the resulting suspension was stirred at room temperature for 10 min. The crystals that precipitated were collected by filtration and triturated with PE/AcOEt /MeOH (3:1:0.2, 5 mL) overnight, filtered and dried at 60 \u003csup\u003eo\u003c/sup\u003eC in vacuo for 2 h to give the target products \u003cb\u003e1a-1e\u003c/b\u003e, \u003cb\u003e1h\u003c/b\u003e, \u003cb\u003e1j\u003c/b\u003e, \u003cb\u003e1m\u003c/b\u003e and \u003cb\u003e1n\u003c/b\u003e. The other crude product (\u003cb\u003e1f\u003c/b\u003e, \u003cb\u003e1g\u003c/b\u003e, \u003cb\u003e1i\u003c/b\u003e, \u003cb\u003e1k\u003c/b\u003e and \u003cb\u003e1l\u003c/b\u003e) was purified by flash chromatography (AcOEt/PE\u0026thinsp;=\u0026thinsp;1:2).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eIn vitro antibacterial assay\u003c/h2\u003e \u003cp\u003eAll strains were grown at 37\u0026deg;C for 12\u0026ndash;16 h in LB until OD\u003csub\u003e600\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.6. Then bacteria were diluted 1:1000 into fresh medium containing TTC, the newly synthesized trifluoromethyl-1,3,4-oxadiazole amide compounds were dissolved in dimethyl sulfoxide (DMSO) to a concentration of 2 mg/mL, and the solutions were diluted with DMSO before antibacterial activity evaluation. For a comparison of antibacterial activity, the commercial gentamicin and ciprofloxacin were used as the reference drugs. Bacteria (198 \u0026micro;L) and compounds (2 \u0026micro;L) were added into the 96 well plates and mixed well by shaking. Then, the plates were incubated at 37\u0026deg;C for 12\u0026ndash;16 h followed by observations of MIC values. For each compound, MIC determinations were performed independently thrice using duplicate samples each time. MIC was determined as the minimum concentration of compound at which visible bacterial growth is inhibited.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eMolecular dock methods\u003c/h2\u003e \u003cp\u003eIn this study, we employed a semi-flexible docking approach to facilitate the formation of stable complexes. Initially, downloaded proteins containing co-crystallized ligands with structures similar to the synthesized compounds from the RCSB Protein Data Bank (PDB ID: 4EI9). The proteins were subsequently prepared using AutoDock Tools 1.5.2 software [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. The preparation phase involved the addition of polar hydrogen atoms and the computation of partial charges. Thereafter, we performed a local charge distribution of the proteins and assigned AutoDock atom types. Subsequently, we established parameters in the Grid module by defining the docking region's box size as 126\u0026times;126\u0026times;126 to encompass the entire protein molecule, thereby aiming to locate the optimal binding site. The grid spacing was set to 0.375 \u0026Aring;, retaining grid information, while other parameters were kept as default [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. The docking process was executed to generate the top ten optimal docking poses. Lastly, visualized the results using PyMol 2.0 software, enabling further analysis of the complex stability and intermolecular interactions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eMolecular dynamics simulation and MMPBSA methods\u003c/h2\u003e \u003cp\u003eTo gain in-depth insights into the intermolecular interactions and stability of the complexes, MD simulations were employed in this study. We simulated the protein-ligand complexes for 100 ns using the GROMACS 2021.3 software package [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. The AMBER99SB-ILDN force field was selected to generate the protein topology files, while the Amber20 software and GAFF force field were utilized to generate the topology files for the small molecule ligands [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. The systems were solvated with a truncated octahedral TIP3P water box extending 10 nm from the molecular surfaces, and Na/Cl ions were added to neutralize the net charge. Energy minimization was carried out using 2500 steps of the steepest descent method followed by 2500 steps of the conjugate gradient method. Subsequently, the systems were equilibrated under NVT conditions for 100 ps at 298.15 K, followed by 100 ps of NPT equilibration [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Finally, 100 ns of production MD simulations were performed under periodic boundary conditions using the NPT ensemble, with long-range electrostatic interactions calculated by the particle mesh Ewald (PME) method. The non-bonded cutoff distance was set to 1 nm, the collision frequency was 2 ps, the system pressure was maintained at 101.325 kPa, the integration time step was 2 fs, and coordinates were saved every 10 ps [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFurthermore, by extracting the stable molecular conformations from the last 20 ns of the equilibrated trajectories, employed the gmx_MMPBSA tool to calculate the binding free energies between the proteins and small molecules [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Through decomposes the binding free energy into individual energy components, such as van der Waals interactions, electrostatic interactions, and polar solvation energies, the relationship between small molecules and proteins was further analyzed from the perspective of energy.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003ch3\u003eAuthor contributions\u003c/h3\u003e\n\u003cp\u003eFei Xiong and Yanjun Zhang were in charge of writing this article and summarizing the data. Yanjun Zhang, Yiren Zhu and Yeji Li performed the synthetic experimental work, and characterization, and molecular dynamic simulation studies. Tianlu Mo and Jinlong Jiao carried out the in vitro antibacterial activity test. Fei Xiong conceived the project and provided the resources, supervision and funding assistance. All authors critically evaluated the manuscript prior to submission.\u003c/p\u003e\n\u003ch3\u003eConflicts of interest\u003c/h3\u003e\n\u003cp\u003eThere are no conflicts of interest to declare.\u003c/p\u003e\n\u003ch3\u003eAcknowledgements\u003c/h3\u003e\n\u003cp\u003eThis work was supported by the financial support of the National Natural Science Foundation of China (NO. 81172918, 51707122), Shanghai Municipal Education Commission (NO. SLG14033) and the open project program of Hubei Key Laboratory of Drug Synthesis and Optimization, Jing Chu University of Technology (NO. OPP2014ZD01). We would like to thank Shiyanjia Lab (www.shiyanjia.com) for the support of NMR tests.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003e\u003cspan\u003eCui P, Li X, Zhu M, Wang B et al (2017) Design, synthesis and antibacterial activities of thiouracil derivatives containing acyl thiourea as SecA inhibitors. 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J Chem Theory Comput 8:3314\u0026ndash;3321\u003c/span\u003e\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Scheme","content":"\u003cp\u003eScheme 1 is available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"molecular-diversity","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"modi","sideBox":"Learn more about [Molecular Diversity](http://link.springer.com/journal/11030)","snPcode":"11030","submissionUrl":"https://submission.nature.com/new-submission/11030/3","title":"Molecular Diversity","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Bacillus cereus, 1,3,4-Oxadiazole, Synthesis, Biological assays, MD Simulation","lastPublishedDoi":"10.21203/rs.3.rs-4148372/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4148372/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThrough the approach of molecular hybridization, this study rationally designed and synthesized new trifluoromethyl-1,3,4-oxadiazole amide derivatives, denoted as \u003cb\u003e1a-1n\u003c/b\u003e. The findings reveal that these novel molecules exhibit potent inhibitory effects against various bacterial strains. Thereinto, compounds \u003cb\u003e1c\u003c/b\u003e, \u003cb\u003e1d\u003c/b\u003e, \u003cb\u003e1i\u003c/b\u003e, \u003cb\u003e1j\u003c/b\u003e and \u003cb\u003e1n\u003c/b\u003e, demonstrate relatively superior antimicrobial performance against \u003cem\u003eB. cereus\u003c/em\u003e FM314, with a minimum inhibitory concentration (MIC) of 0.03907 \u0026micro;g/mL. Molecular docking analysis suggests the potential importance of the Ser57 and Thr125 amino acid residues in contributing to the inhibitory activity against \u003cem\u003eB. cereus\u003c/em\u003e. The consistency of these results was further corroborated through subsequent molecular dynamics simulations and MMPBSA validations. The insights gained from this study serve to facilitate the rational design and efficient development of novel eco-friendly antimicrobial inhibitors based on the trifluoromethyl-1,3,4-oxadiazole amide scaffold.\u003c/p\u003e","manuscriptTitle":"Towards new bioactive fluorine-containing 1,3,4-oxadiazole-amide derivatives: Synthesis, antibacterial activity, molecular docking and molecular dynamics simulation study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-28 02:02:54","doi":"10.21203/rs.3.rs-4148372/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-04-11T04:08:13+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-04-06T12:08:28+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-03-27T13:03:15+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"76891cda-7e22-4787-bc21-d729771e34dd","date":"2024-03-26T02:53:22+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"f537a083-3ebb-458b-87b3-be4f16b14e9b","date":"2024-03-26T02:23:34+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-03-26T02:00:09+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-03-22T11:03:19+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-03-22T11:01:35+00:00","index":"","fulltext":""},{"type":"submitted","content":"Molecular Diversity","date":"2024-03-22T08:53:36+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"molecular-diversity","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"modi","sideBox":"Learn more about [Molecular Diversity](http://link.springer.com/journal/11030)","snPcode":"11030","submissionUrl":"https://submission.nature.com/new-submission/11030/3","title":"Molecular Diversity","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"1bfad2c4-8267-43d6-baf8-d81f67128861","owner":[],"postedDate":"March 28th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-06-21T16:10:42+00:00","versionOfRecord":{"articleIdentity":"rs-4148372","link":"https://doi.org/10.1007/s11030-024-10893-x","journal":{"identity":"molecular-diversity","isVorOnly":false,"title":"Molecular Diversity"},"publishedOn":"2024-06-20 16:10:42","publishedOnDateReadable":"June 20th, 2024"},"versionCreatedAt":"2024-03-28 02:02:54","video":"","vorDoi":"10.1007/s11030-024-10893-x","vorDoiUrl":"https://doi.org/10.1007/s11030-024-10893-x","workflowStages":[]},"version":"v1","identity":"rs-4148372","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4148372","identity":"rs-4148372","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}