Quinolinone Schiff bases and fused pyridazinoquinolinone derivatives represent canonical classes of anticancer and broad biological profiles. Herein, a panel of parent quinoline scaffolds clubbed with endowed nitrogenous entities involving amines, hydrazines, and pyridazines was designed and synthesized through direct methodologies. The established scaffolds inculpating different moieties such as benzene, benzo[d][1,3]dioxole, quinoline, chromone and ferrocene as bioactive hybrids for obtaining promising surrogates targeting anti-lung cancer and anti-tuberculosis through strategies for drug optimization Fig2.
The condensation of 3-acetyl-4-hydroxy-1-phenylquinolin-2(1H)-one (AHQ) (1) with functionalized mono /di-nucleophilic amino derivatives such as m-anisidine, o-phenylenediamine (OPD), and p-phenylenediamine (PPD) afforded the corresponding Schiff bases 2, 4, and 5, respectively (Scheme 1). Whereby, the ortho position for 3-methoxyphenylimino substrate of Schiff base 2 is strongly activated toward SAr, resulting in a preference for 6-membered ring cyclization with the pendant hydroxy group of quinolinone under acidic conditions. However, the attempts for formation of compound 3 was unsuccessful either direct or indirect reactions (Scheme 1). The chemical structures for the Schiff bases 2, 4, and 5 were elucidated by a combination of H, C NMR, mass spectrometry (MS), and IR techniques that are in accordance with their suggested structures. For constitution 2, its IR spectrum displayed bands at 1640 and 3420 cm attributed to amidic carbonyl and OH groups, respectively. Whereas, its H NMR spectrum shows two singlet signals at δ 2.64 and 3.78 ppm corresponding to the methyl and methoxy groups and a downfield signal at 16.53 ppm for the OH group. On the other hand, the IR spectrum of structure 4 reveals two bands at 3421 and 3336 cm for the NH group besides the lactamic band at 1634 cm. Whereby, the H NMR spectrum for scaffold 4 revealed three singlet signals at δ 2.55, 5.31, and 15.82 ppm for CH, NH and OH. Also, the MS spectrum of 4 showed a molecular ion peak at m/z 369.53, compatible with its molecular formula CHNO. Similarly, aiming to synthesize poly-functionalized bis-quinolinone via reaction of AHQ 1 with PPD furnished (E)-3-(1-((4-aminophenyl)imino)ethyl)-4-hydroxy-1-phenylquinolin-2(1H)-one 5 instead of the desired bis product. Its H NMR chart displays singlet signals at δ 2.63, 5.45, and 15.84 ppm for CH, NH and OH groups, respectively. Interestingly, the inserted aromatic protons of PPD appeared as two doublet signals at δ 6.65 and 7.04 for C3-C2 with J= 8.4 Hz.
The scope of this tandem route was extended by the reaction of AHQ 1 with tryptamine at pH 4.7 (physiological conditions) according to the Pictet-Spengler reaction furnished (E)-3-(1-((2-(1H-indol-3-yl)ethyl)imino)ethyl)-4-hydroxy-1-phenylquinolin-2(1H)-one 6 instead of the desired product tetrahydro-β-carboline system 7. Various spectral data established the molecular structure of 6, as its H NMR spectrum revealed singlet signal at δ 2.65 ppm for methyl group, two signals at δ 3.22, 3.84 ppm characteristic for two methylene groups, singlet signal at 8.16 ppm characteristic for proton of indole and broad signal at δ 15.06 ppm for OH group. The C NMR spectrum displayed new signals at δ 25.4, and 44.6 ppm for two methylene groups. The mass spectrometry measurement of compound 6 shows a molecular ion peak at m/z 421.74, which confirms the recommended structure (Scheme 1).
The stupendous chemical reactivity of 1 toward binucleophiles was investigated through the treatment of AHQ 1 with hydrazine hydrate in EtOH or CHCl afforded (E)-3-(1-hydrazineylideneethyl)-4-hydroxy-1-phenylquinolin-2(1H)-one (8). It is worth mentioning that this reaction had previously been carried out in the presence of EtOH as the solvent; however, it required 24 hours to complete the reaction Therefore, a fundamental modification was made to the preparation methodology to obtain a pure sol product within 1h at ambient temperature by using CHCl as a weak polar solvent as an alternative solvent (Scheme 2).
The structure of compound 8 was elucidated by various spectral analyses, as its H NMR spectrum displays the presence of a singlet signal at 2.62 ppm characteristic of a methyl group alongside the presence of a characteristic singlet signal at δ 6.11 ppm attributed to NH. The MS spectrum exhibits its molecular ion peak at m/z 293.28, which is attributable to the suggested structure.
In a similar manner, the chemical reactivity of 1 towards various arylhydrazines was implemented through the treatment of 1 with phenyhydrazine, 1-methyl-1-phenylhydrazine, and 2,4,6-trichlorophenylhydrazine in EtOH under stirring conditions at room temperature for 2h, furnished hydrazones 9a-c (Scheme 2). The constitution of compound 9b was confirmed by its H NMR spectrum, displaying the presence of two singlet signals of two methyl groups at δ 2.82 and 3.25 ppm and a singlet signal attributed to OH at δ 15.42 ppm. Also, C NMR spectrum assured the presence of signals at upfield δ 17.0 and 41.7 ppm for two methyl groups, besides downfield signals at δ 179.4, 177.4, and 163.5 attributed to (C-OH), (C=O, lactamic), and (C=N) groups. The mass spectrum confirms the constitution of scaffold 9b, showing its molecular ion peak at m/z 383.54 as compatible with the suggested structure. The H NMR spectrum of hydrazone 9c displays the appearance of three singlet signals at δ 2.80, 8.44, and 15.58 ppm characteristic of CH, NH, and OH functionalities. Whereby, its IR spectrum showed two characteristic bands at ν1640 and 3241 cm related to the lactamic carbonyl and NH groups, respectively.
Due to the renowned biological activities of the quinolinone and pyrazole moieties, a combination of quinolinone and pyrazole in one molecular framework is one of the main targets for this manuscript. Subsequently, hydrazone 9a was subjected to react with Vilsmeier-Haack reagent (DMF-POCl) to afford binary pyrazoloquinolinone scaffold 10 (Scheme 2). The spectral analyses of compound 10 were in accordance with the suggested structure, as its H NMR shows singlet signals at 9.34 and 9.83 ppm characteristic of HC and formyl protons, respectively. Whereas aromatic protons appeared as a multiplet at δ 7.55-7.65 ppm. On the other hand, its C spectrum displays a characteristic signal at δ 184.9 ppm corresponding to the formyl group, and upfield signals of aliphatic carbons are absent. Whereby, the mass spectrometry of scaffold 10 exhibited its molecular ion peak at m/z 425.14 and M+2 at m/z 427.12 corresponding to chlorine isotopes, which is attributable to the suggested structure.
The synthetic route for the binary quinolinone-pyrazole 10 is rationalized through an elegant mechanistic pathway that involves the generation of an iminium salt as a Vilsmeier-Haack reagent. Next, the terminal methyl electron-rich attacks the iminium salt, leading to intermediate 10A. Whereby, nucleophilic attack of secondary NH functionality followed by cyclization and subsequent elimination of chloride ion to generate intermediate 10B. Further, the intermediate 10B encounters another iminium salt, which leads to the formation of intermediate 10C. Then, the dimethylamine molecule is stripped off to yield intermediate 10D. After that, the hydrolysis process of intermediate 10D results in the formation of binary polycyclic formyl pyrazole intermediate 10E. Finally, enolizable hydroxy proton is subjected to the chlorination process due to the excess effect of the POCl reagent afforded dihydroquinolin-1-phenyl-1H-pyrazole-4-carbaldehyde 10 (Scheme 3)
Whereas, stirring of AHQ 1 with phenylhydrazine in CHCl at room temperature for 2h produced 3-methyl-1,9-diphenylpyrazolo[3,4-b]quinolinediol 11 instead of the anticipated Schiff base 9a (Scheme 2). This reaction proceeds via a condensation reaction followed by nucleophilic attack of NH functionality to the electrophilic lactamic carbonyl group, followed by ring closure to yield fused pyrazolo[3,4-b]quinoline 11. The proposed structure 11 was elucidated by mass spectrometry measurement that showed a molecular ion peak at m/z 369.46. H NMR spectrum assured the presence of three singlet signals at δ 2.64, 9.55 and 16.23 ppm for the methyl group and two exchangeable OH groups.
Whereby, the difference in reactivity related to the utilized solvents was observed by Hamama et al. reported the chemoselectivity of the true carbonyl group through using a highly polar solvent (EtOH), whereas the use of a less polar solvent (CHCl) led to the localization of the electropositivity on the other less reactive carbonyl group, and the viability of these postulations was supported by density functional theory (DFT) calculations.
Analogously, the most acceptable explanation for formation of compound 11 was recognized through calculating the electron density values via density function studies (DFT) using BY3LP as function of charge on each atom, as the using of the CHCl as solvent (less polar solvent) alters the logic charge distributions of compound 11, leading to accumulation of negative charge on NH functionality makes it more nucleophilic position (-0.453), whereby the most electropositive charge is allocated on the lactamic carbonyl group (+0.472) than other enolic carbon at position 4 (+0.422) as presented in figure 3.
Additionally, reaction of 1 with phenylhydrazine hydrochloride in a refluxing mixture solvent system of glacial acetic acid and concentrated hydrochloric acid (4:1) according to the Fischer indole synthesis procedures afforded 3-methyl-1,5-diphenyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one (12) in 88% yield instead of the anticipated spiro Fischer indole product 12. The structure of compound 12 was confirmed by its H NMR spectrum, which displayed the presence of a singlet signal of methyl groups at δ 2.56 ppm. The C NMR spectrum showed a signal at δ 12.8 ppm for the methyl carbon. Whereas, the IR spectrum showed a broad band at 1667 cm characteristic of lactamic group. The mass spectrum of this compound showed a molecular ion peak at m/z 351.61 (35.43) and 352.27 (M+1, 26.97), confirming the proposed structure.
Isoniazid (INH) is a trademarked antibiotic used to treat both latent and active tuberculosis, besides demonstrating potential anti-proliferative actions. Till now, INH is still considered the first-line drug against TB due to its lesser toxicity, higher efficacy, and high aqua-solubility. Furthermore, it causes inhibition of the synthesis of mycolic acid as an essential component of the mycobacterial cell wall Based on the merits as mentioned above, and in continuation with our trials to synthesize and develop new anti-cancer agents, INH was utilized to react with AHQ 1, affording 1,2-dihydroquinolin-3-yl(ethylidene)isonicotinohydrazide 13 with an unsuccessful trial of its formylation through the Vilsmeier-Haack reaction (Scheme 2). The constitution of hydrazone 13 was established based on IR, H NMR, and mass spectra. As its IR spectrum shows bands at 1615 and 1688 cm corresponding to two carbonyl groups, and the appearance of a strong band at 3201 cm is characteristic of NH. Also, its H NMR spectrum displays the presence of three singlet signals at δ 2.72, 11.96 and 16.88 ppm attributed to the methyl, NH and OH groups, respectively. Whereas the MS spectrum of compound 13 exhibited its molecular ion peak at m/z 398.02, which confirms the suggested structure.
Pyridazine derivatives are aromatic heterocyclic compounds belonging to the diazine
family, characterized by nitrogen atoms at the 1,2-positions. Also, pyridazine can be utilized as a bioisostere for benzene or pyridine. Compared to other diazines, pyridazines exhibit higher pKa values (indicating greater basicity) and higher dipole moments. Besides, pyridazines have high potential for coordination with metals and forming hydrogen bonding, as well as their polarity increases due to the presence of additional nitrogenous atoms, leading to the formation of more water-soluble salts or crystals Thus, the reaction of hydrazone derivative 8 with aldehydes seemed to be a unique route for synthesizing promising annulated pyridazinoquinolinone systems according to ring closure Baldwin's rules via 6-exo-trig cyclization (Scheme 4).
The mechanistic pathway of the reaction of skeleton 8 and various aldehydes is described in Scheme 5, starting with a condensation reaction giving dihydrazone intermediate that isomerizes to ketoform, followed by 6-exo-trig cyclization yielding six new-fangled pyridazine entities 14-19 (Scheme 5). Consequently, treatment of hydrazone 8 with piperonal yielded 1-(benzo[d][1,3]dioxol-5-yl)-4-methyl-6-phenylpyridazino[4,5-c]quinolin-5(6H)-one (14) that was elucidated by different spectroscopic analyses, which were consistent with its proposed structure. As its H NMR spectrum demonstrates a singlet signal of the methyl protons at δ 3.10 ppm, it also shows a singlet signal at δ 6.04 for the methylene group of the piperonal moiety and has demonstrated the disappearance of NH and OH protons related to the parent structure of constitution 8. Whereby, its IR spectrum shows a band at 1643 cm characteristic of the lactamic C=O group. The mass spectrometry showed a molecular ion peak at m/z 407.93 (M, 18.39) corresponding to a molecular formula CHNO.
Similarly, cyclocondensation reactions of hydrazone 8 with 2-chloroquinoline-3-carbaldehyde, 4-oxo-4H-chromene-3-carbaldehyde, and ferrocenecarboxaldehyde were carried out affording pyridazino[4,5-c]quinolin-5(6H)-one derivatives 15-17, respectively. As the IR spectra of products 15-17 show the presence of characteristic lactamic peaks at 1625-1643 cm due to highly conjugated systems.
The H NMR spectrum of compound 16 reveals a singlet signal at δ 3.00 ppm for CH and a sharp singlet signal at δ 9.07 ppm for the olefinic proton of the chromone moiety. Its C NMR spectrum demonstrates deshield signals at δ 174.8 and 158.0 ppm corresponding to carbonyl groups of C=O and lactamic groups. Whereas, H NMR spectrum for 17 gives singlet signals at δ 4.29, 4.58, and 4.75 ppm characteristic for protons of ferrocene moiety, the MS spectrum of 17 shows a molecular ion peak at m/z 471.92 (M, 27.34) coinciding with a molecular formula CHFeNO.
In the course of this study, the synthesis of bis-pyridazino[4,5-c]quinoline entities was investigated through the reaction of hydrazone 8 with glyoxal and terephthaldehyde as dialdehyde constitutions afforded the bis-cyclized product bipyridazino[4,5-c]quinolin]-5,5'(1H,6'H)-dione 18 and 6-phenyl-5,6-dihydropyridazino[4,5-c]quinolin-1-ylbenzaldehyde 19, respectively. These structures are proven by various elemental analyses and spectral data. As the H NMR spectrum of 18 reveals a singlet signal at δ = 3.03 ppm for two symmetrical methyl groups and the absence of any signal related to formyl groups, confirming that the condensation reaction occurred at the two formyl groups of glyoxal. Moreover, C NMR spectrum showed signals at δ 181.4 and 174.6 ppm for two lactamic C=O, and showed two signals at δ 17.6 and 17.4 ppm for two methyl groups. Finally, the mass spectrometry measurement of 18 shows a molecular ion peak at m/z 572.00 (M, 14.50), confirming the proposed structure. On the other hand, the H NMR spectrum of compound 19 demonstrates two singlet signals at δ 3.15 and 10.07 ppm attributed to methyl protons and the formyl proton. Meanwhile, characteristic signals were seen in the C-NMR spectrum for 19 at δ 191.4 and 180.9 ppm corresponding to two carbonyl groups. Whereas, the MS spectrum provides robust evidence to the suggested constitution displaying a molecular ion peak at m/z 391.65 (M, 27.61) and a base peak at m/z 63.20, verifying a molecular formula CHNO.
The success of the cascade cyclization methodology encouraged us to repeat this reaction using a ketonic scaffold, aiming at extending the scope of our strategy through treatment of hydrazone 8 with different ketonic compounds, which afforded the opened condensation products 20-22, not the cyclized one 23. Thus, the reaction of 8 with acetophenone, 6-acetyltetralin and DHA yielded dihydrazone 20-22 (Scheme 6). The structures of hydrazones 20-22 were unambiguously elucidated using various spectroscopic analyses. As H NMR spectrum for 20 demonstrated two singlet signals of the two methyl protons at δ 2.63 and 3.14 ppm, its C NMR spectrum supports the suggested structure as recording sixteen signals corresponding to the diverse carbon atoms of the molecular formula CHNO. In addition, the EI-MS spectrum revealed its molecular ion peak at m/z 395.10, which was in accordance with its molecular formula.
Whereby, EI-MS reinforced the constitution of compound 21 due to the appearance of its molecular ion peak at m/z 449.28 (M, 16.18), which was compatible with its constitution. Also, H NMR spectrum of designated 21 confirms the presence of upfield signals in the range of δ 1.75 and 2.78 ppm related to aliphatic protons of the tetrahydronaphthalenyl moiety. Whereas, H NMR spectrum of 22 shows three singlet signals at δ 2.22, 2.88, and 2.95 ppm related to three methyl groups, alongside the presence of two exchangeable singlet signals at δ 16.07 and 17.30 ppm characteristic for two hydroxyl groups, whereby their carbons resonated at δ 182.3 and 164.4 ppm in C NMR analysis. Furthermore, the IR spectrum showed two characteristic carbonyl signals related to lactonic and lactamic carbonyl at 1717 and 1644 cm, respectively. Its mass spectrum revealed a peak at m/z 443.39 corresponding to its molecular ion.
According to Baldwin's rules, an undesired (slow) reaction does not have a rate capable of competing effectively with the preferred (fast) alternative reaction. Additionally, DFT studies confirmed the obtained product 20a, which predominates over the others (20b,20c) due to chemical stability (lower total energy, -34831.312 eV) than the others (-34820.768 and -34828.863 eV, Fig. 4.
Biological evaluation
All the target compounds were screened for their potential in vitro anticancer activity against the lung cell strain (A549 cell line, which was obtained from VACSERA, Cairo, Egypt) by MTT assay. Also, they were tested for their potential in vitro antituberculosis activity against the Mtb H37Rv strain by Microplate Alamar Blue Assay (MABA).
In vitro anticancer screening
Initially, all of the newly synthesized quinolinone derivatives and the starting materials were examined in vitro to quantify their inhibitory activity against the anticancer (A549 cell line). Anticancer was performed using the MTT (Thiazolyl Blue Tetrazolium Bromide) method Doxorubicin, one of the common anti-cancer drugs, was employed for comparison. The concentration of compounds needed to inhibit the growth of 50% of cancer cells was expressed as (μM) and displayed in Table 1. The compounds under examination demonstrated varying degrees of inhibitory effects on the tested human tumor cells. All synthesized compounds and the starting material were evaluated for their efficacy in inhibiting lung cancer compared to Doxorubicin (IC= 4.18 µM). The inhibition potency of the synthesized compounds was in the range of 10.38-over 100 µM. Compound 15 exhibited remarkable activity with an IC value of 10.38 µM. Further, the compounds 10, 14, 16, 19, and 21, are proven to be of distinguished activity with IC values of 25.23, 30.55, 12.40, 15.91, and 18.01 µM, respectively. On the other hand, compounds 9b, 9c, 12, 17, 20 and 21 demonstrated moderate activity with IC values of 52.32, 57.62, 35.31, 54.12, 34.54 and 59.52 µM, respectively. The derivatives 2, 4, 5, 6, 8 and 11 recorded weak values of IC 71.54, 77.33, 80.23, 73.65, 92.22 and 93.23 µM, respectively. Finally, compounds 1, 13, and 18 showed no acceptable growth inhibitor effect (IC >100) (Table 1). Pyridazino[4,5-c]quinoline derivatives (14-19) demonstrated the highest activity with IC in the 10.38-30.55µM range. Figure 5 shows the A549 cell line viability that is highly sensitive to compounds 15, 16, and 19 in the presence of varying concentrations.
In vitro antituberculosis screening
Next, all the newly synthesized compounds were initially screened for their in vitro tuberculosis activity at a concentration of 3.125 mg/mL against Mtb H37Rv strain using the MABA. Isoniazid, one of the common antituberculosis drugs, was employed for comparison with compounds. The results of the antitubercular studies are presented in Table 2. The compounds under examination demonstrated varying degrees of inhibitory effects on Mtb (H37Rv) strain. All synthesized compounds and the starting material were evaluated for their efficacy in inhibiting Mtb (H37Rv) strain in comparison to Isoniazid (MIC= 3.12 µM). The inhibition potency of the synthesized compounds was in the range of 6.25-32.26 µM. Compounds 15 and 14 exhibited remarkable activities with an MIC value of 6.25 and 8.37 µM, respectively. Further, the compounds 6, 16, 19 and 21 are proven to be of high activity with MIC values of 18.58, 11.34, 12.25 and 14.72 µM, respectively. On the other hand, compounds 4, 5, 9b, 9c, 10, 11, 12, 13, 20, and 22 demonstrated moderate activity with MIC values ≤30 µM. The derivatives 2, 8, and 18 recorded values of MIC >30 µM. Finally, compound 1 (starting material) showed the lowest value of MIC (Table 2).
Molecular simulations
Major efforts have been devoted to improving the algorithm for docking predictions, as molecular docking is an essential tool for rational medication design in the disciplines of biology and pharmacology. In light of that, molecular simulations can be used to analyze the potential interactions between produced drugs and protein receptors, providing vital insights into their binding patterns and potential antituberculosis and anticancer activity This analysis was intended to determine the efficacy of the synthesized scaffolds. The drug-target interactions were examined using molecular docking analysis to clarify the binding sites and to understand the mode of action for the high-potential synthesized candidates. Compounds 6, 10, 14, 15, 16, 19, 20, and 21 were selected based on their high biological activity, indicating higher binding energies and higher therapeutic efficacy. The highest binding energy conformation was considered the most favorable docking pose. The binding energies of these compounds are presented in Tables 3 and 5. The components are as follows: (1) Final Intermolecular Energy, including Van der Waals forces, hydrogen bonds, and desolvation energy; (2) Final Total Internal Energy; (3) Torsional Free Energy; and (4) Energy of the Unbound System. The formula [(1) + (2) + (3) - (4)] was used to calculate the binding energies for each derivative in kcal/mol. Moreover, the K value or the inhibition constant is in fact the dissociation constant of the docked enzyme inhibitor complex. The smaller the value of K, the lower will be the probability of dissociation, and hence higher will be the inhibition. It is calculated as Ki = exp(ΔG/(R*T)) where ΔG is the free energy of binding, R is the gas constant (1.987 cal K mol), and T is the temperature (298.15 K).
Whereas, epidermal growth factor receptors (EGFRs) are transmembrane receptors present on cell membranes, represent an effective role in controlling cellular functions involving cell growth, cell proliferation and apoptosis. In addition, the mutations of EGFRs can lead to abnormal stimulation of the receptors causing unregulated cell division, causing cancer such as NSCLC Whiles, 1M17 is a reprehensive protein code for EGFR tyrosine kinase domain as a crucial element in cell signaling related to growth and proliferation as a well-established anticancer inhibitor On observing the docking score of the most bioactive compounds with 1M17, it was observed that 15 displayed the higher docking score (-9.97 kcal/mol) followed by 16, 19, 21, 10 and 14 with binding energy -9.89, -9.59, -9.37, -9.08 and -8.84 kcal/mol (decreasing order), compounds with higher docking score are pyridazino[4,5-c]quinolinone hybrids due their biological activity as illustrated in Table 3. The 2D and 3D docking interaction pattern revealed that essential interactions were present in all docked synthesized compounds, as it is clearly visible from Figure 6 and Table 4.
On the other hand, 1OUZ is a vital code for the InhA protein which plays a crucial role in the development of antitubercular drugs, as it is an effective carrier protein reductase from Mycobacterium tuberculosis, so inhibiting process for this enzyme prevents the bacterium from synthesizing its cell wall that is essential for the pathogen's viability and survival. The docking score of the highly active compounds with 1OUZ (M.tb protein) supported the biological activity of the synthesized compounds. Also, it was observed that 15 displayed the highest docking score (-10.59 kcal/mol) followed by 14, 16, 19, 21 and 6 with binding energies of -10.12, -10.05, -9.79, -9.51 and -9.41 kcal/mol (decreasing order) (Table 5). The 2D docking interaction pattern revealed that essential interactions were present in all docked synthesized compounds. Moreover, it is clearly visible from Figure 7 and Table 6.
To correlate the structure with biological activity, studying some 3D-descriptors, such as HOMO and LUMO frontier molecular orbitals, was interesting. The chemical reactivity was reflected through the energy band gap (E-E) as an important stability index and influences the biological activities of the molecules, global hardness (η), global softness (S), electronegativity (χ), global electrophilicity index (ω), and nucleophilicity index (ε), that were calculated from energies of frontier molecular orbitals (E, E), by Equations (1-6) as follow Fig.8, 9:
Compounds 14, 15, 16, and 19 showed high potency in both tested biological applications with closely comparable softness values (0.48-0.54). Based on the electronic descriptors, compound 10 exhibits a lower energy gap (3.37 eV) and a high softness value. Theoretically, the presence of binary pyrazole functionality will impart a strong activity, yet it showed moderate activity compared to other tested scaffolds in the practical part. This notification may be attributed to the solubility of the compound and its lipophilicity and permeability across the lipid bilayer membrane of the cell. Whereby, the formyl, lactamic functions plus the two adjacent nitrogenous atoms in the pyrazole ring led to increasing the polar surface area (PSA); reducing the lipophilicity and thus its capability to passively diffuse over the non-polar cell membrane in contrast to the targeted drug-like profile (low water solubility and high lipophilicity)