Synthesis and Biological Evaluation of Some New 3-Aryl-2-thioxo-2,3-dihydroquinazolin-4(1H)-ones and 3-Aryl-2-(benzylthio)quinazolin-4(3H)-ones as Antioxidants; COX-2, LDHA, α-Glucosidase and α-Amylase Inhibitors; and Anti-Colon Carcinoma and Apoptosis-Inducing Agents

1. Introduction

Colorectal cancer (CRC) was ranked third and second in terms of incidence and mortality, respectively, accounting for 1.9 million of all new cancer cases and 900,000 deaths in 2020 worldwide. Notably, the incidence of CRC has shown a rising trend among individuals aged less than 50 years [1].

CRC neo-tumorigenesis is a complex and multistep process, linked to factors of oncogenesis such as oxidative stress (OS), chronic inflammation, reprogrammed glucose metabolism and diabetes mellitus (DM). In addition, defective signaling or changes in the expression of proteins regulating programmed cell death ‘’apoptosis’’ play crucial roles in the evolution of normal mucosa cells towards neoplastic transformations and the acquisition of new characteristics, among which evading cell death [2].

OS is a pathological state featuring the over-generation of cellular reactive oxygen and/or nitrogen species (RO/N S) as a result of a dysregulated intracellular metabolism, tissue inflammation or exposure to exogenous stimuli at an extent that exceeds the cellular antioxidant defenses [3]. In fact, OS is considered as a hallmark of carcinogenesis [3].

Regarding CRC, OS on intestinal mucosal cells is proven to damage phospholipid biological membranes via lipid peroxidation by free radicals [4]. Therefore, it causes rupture of the phospholipid bilayer of the cytoplasmic cellular membrane, with the loss of cellular homeostasis. It promotes the destruction of the two lipid bilayers of the endoplasmic reticulum (ER), the dynamic reservoir of Ca²⁺, leading to disruption of ER-Ca²⁺ homeostasis, which is implicated in the initiation and propagation of cancer [5]. Also, it causes oxidative damage to the mitochondrial membranes, which disrupts the cell energy metabolism [6].

Another aspect of involvement of OS in CRC pathogenesis is through production of malondialdehyde (MDA, one of the end products of lipid peroxidation), which promotes oxidative DNA damage via formation of adducts with DNA bases; deoxyguanosine and deoxyadenosine. s, thus initiating mutations in tumor suppressor genes and proto-oncogenes [6,7].

Furthermore, free radicals may modify proteins, thus diminishing or enhancing their catalytic activity. Also, free radicals may stimulate cell and tissue lesions by oxidizing carbohydrates [8], and they may exert pro-inflammatory and pro-proliferative roles via oxidizing cholesterol to oxysterols [9]. Interestingly, cancerous cells can maintain certain levels of ROS to promote their proliferation and invasion [4]. Moreover, ROS are key constituents of cancer cells’ survival via the generation of resistance to chemo- and radiotherapies [10]. Accordingly, several studies have indicated that antioxidant compounds were supplemented or tested as adjuvants in cancer therapy to reverse resistance mechanisms, to reduce systemic toxicity during chemo- or radiotherapy [11] and to prevent cancer cell invasion [12].

Chronic inflammation is another principal effector in colorectal carcinogenesis. Inflammation can be induced by different stimuli, including pathogens, toxic compounds and OS-damaged cells. This inflammation enhances the affected tissues to overexpress the inducible cyclooxygenase isoform-2 (COX-2), which is typically absent or rarely expressed in healthy tissues. COX-2 catalyzes the conversion of arachidonic acid (AA) to prostaglandins (PGs) H₂, which are further metabolized by prostaglandin synthases to PGD₂, E₂, I₂, F_2α and thromboxane A₂. Extensive clinical and experimental studies conducted over the last few decades have provided convincing data indicating that high levels of COX-2 and PGE₂ are implicated in the pathogenesis of colorectal carcinoma [13,14,15], and that they are associated with poorer survival outcomes [16]. The key roles of the COX-2/PGE2 pathway in CRC pathogenesis include generating an immunosuppressive tumor microenvironment [17], participating in tumor initiation, promoting sustained angiogenesis and thus favoring tumor growth, inducing tumor cell invasion, encouraging migration, reducing apoptotic rate and enhancing chemotherapeutic resistance [18,19]. Therefore, inhibition of COX-2 to reduce PGE2 synthesis is considered an important therapeutic approach in prevention and modulation of CRC progression [20]. Consistent with these findings, a number of meta-analyses and in vitro studies have reported that classical non-steroidal anti-inflammatory drugs (NSAIDs) [21] and selective COX-2 inhibitors [22] provided protection against CRC incidence, reduced mortality and improved chemosensitivity and the efficacy of anticancer drugs when co-administrated [23].

Furthermore, the reprogrammed energy metabolism or Warburg effect [24] is a prominent metabolic characteristic of malignant cells [25]. The common features of this abnormal, less-energy-efficient metabolic pathway include increased glucose uptake, production of two moles of pyruvate and less ATP (2 moles/ glucose molecule) and rapid cytoplasmic fermentation of pyruvate to lactate under catalysis of lactate dehydrogenase A (LDHA). Lactate is then exported to the extracellular environment, which, in the presence of hydrogen ion, generate an acidic tumor microenvironment (TME). Lactic acidosis has been observed in different cancer types and it is linked to tumor aggressiveness [26]. In this regard, several clinical studies have revealed that hyperlactatemia is involved not only in tumor growth but also in the evasion of immune surveillance [27], local invasion, metastasis, chemoresistance and patient survival [28]. Consequently, targeting glycolysis via inhibiting LDHA would suppress hyperlactatemia and prevent its protumorigenic effects, in addition to depriving cancerous cells of nutrients, thereby killing them [29]. Indeed, several studies have shown that inhibition of LDHA activity impedes tumor progression, induces significant oxidative stress and necrosis [30], induces G2/M cell cycle arrest, increases sensitivity to ionizing radiation and 5-Fluorouracil (5-FU) [31,32] and attenuates cell invasion and migration [33].

DM is a multifactorial chronic metabolic disorder affecting millions of people worldwide. DM is categorized into Type 1 (T1DM, caused by the autoimmune destruction of the pancreatic β-cells leading to little, or the absolute lack of, insulin production), Type 2 (T2DM, arises due to complex interactions between genes and some risk factors, mainly obesity, smoking and hypertension, leading to insulin resistance) and gestational (GDM, occurs in pregnant women). T2DM is the most prevalent type, accounting for about 90% of all diabetic cases [34]. Besides its deadly cardiovascular complications, T2DM has also been associated with an increased risk for the development of certain types of cancers [35]. In particular, the positive correlation between CRC and T2DM is attributed to sharing some common risk factors including obesity, alcohol consumption, cigarette smoking and a Western-pattern diet. In addition, genome-wide association studies have identified transcription factor 7-like 2 (TCF7L2), gremlin 1 (GREM1) and tumor protein p53 inducible nuclear protein 1 (TP53INP1), among other genes, as being pleiotropically related to T2DM as well as CRC prognosis and tumorigenesis [36]. Moreover, T2DM-related metabolic changes, such as hyperinsulinemia, hyperglycemia, OS, inflammation induced by adipose tissue dysfunction, impaired immunological surveillance and gastrointestinal motility disorder have been implicated in carcinogenesis [37].

With regard to hyperglycemia, it influences the neoplastic transformations by providing glucose that is essential for the growth and proliferation of cancerous cells under the Warburg state [38]. It leads to the irreversible formation of advanced glycated end-products (AGEs) with proteins, lipids and nucleic acids, as well as their buildup, resulting in the development of OS and chronic inflammations, which are intrinsically linked to induction and development of cancer. Furthermore, elevated expressions of AGEs and their receptors contribute to the invasion and metastasis of CRC [39]. Also, the activations of the polyol, hexosamine and protein kinase C metabolic pathways [40] are other putative mechanisms by which hyperglycemia contributes to the cell proliferation, invasion and tumor progression of colon cancer. In addition, hyperglycemia may negatively affect CRC patients’ quality of life by reducing the chemotherapeutic efficacy and inducing adverse effects [36].

Thus, over the years, researchers have worked on various targets to moderate the onset or worsening of diabetes, from which some antihyperglycemic therapies have been approved by the US FDA and other regulatory agencies, which exhibit different mechanisms of action via targeting single or multiple established molecular pathways involved in the pathogenesis of the disease [41]. Amongst these validated pathways is the breakdown of complex carbohydrates into absorbable monosaccharides, which takes place under the catalysis of alpha-glucosidase (AG) and alpha-amylase (AA). Thus, inhibitors (Is) of these enzymes would manage T2DM by delaying the carbohydrate metabolism and turning down the rate of glucose absorption to reduce the post postprandial blood glucose level (PPBGL). Indeed, carbohydrate mimics [34], including voglibose (natural sugar), miglitol (semisynthetic iminosugar) and 1-deoxynojirimycin (DNJ), all of which inhibit AG, in addition to acarbose (an oligosaccharide), which can also inhibit AA [42], are marketed as antidiabetic drugs. Despite the helpful impacts of these glucose-lowering drugs, morbidity and mortality remain significant in T2DM patients. In addition, these medications have many drawbacks, such as poor compliance with treatment, expense and adverse effects [43,44]. Therefore, there are a lot of synthetic efforts towards the development of new inhibitors, particularly annulated sugars [45], iminosugars [46,47] and novel classes of drugs derived from the structural hybrids of different key molecules and heterocycles [48], with the aim to develop new effective inhibitors that are escorted by better compliance, socioeconomic benefits and safety [49].

Additionally, apoptosis is a well-characterized form of programmed cell death (PCD), which occurs normally for regulating tissue homeostasis and regeneration during development. However, it is also activated as an immune defense mechanism against damaged or stressed cells in response to stimuli such as reactive oxygen species (produced by some chemotherapeutic agents) or DNA damage (from irradiation or anticancer drugs) to remove these potentially harmful cells [50]. If apoptosis in not controlled properly (insufficient apoptosis/extreme cell proliferation), mutations will potentially accumulate in the unwanted cells, which eventually could lead to the induction of cancer. Indeed, resisting apoptosis is considered as one of the hallmarks of cancer [51].

Apoptosis can be triggered by two different pathways: the extrinsic (death receptor) and the intrinsic (mitochondrial) pathways. The extrinsic pathway is initiated via the binding of certain extracellular ligands (members of the tumor necrosis factor super family, TNF) to their cognate cell surface death receptors, resulting in caspase 8 activation. This activation can directly induce apoptosis, or activate effector caspase 3 or Bid, which lead to apoptosis. While the intrinsic apoptotic pathway is initiated via mitochondrial outer membrane permeabilization (MOMP), a process controlled by the members of the B-cell lymphoma (BCL-2) family and p53 proteins [52]. MOMP is considered as the point of no return in this apoptotic cascade, since it leads to the release of the apoptogenic proteins cytochrome c and Smac from the mitochondria into the cell cytoplasm, which activate caspases directly or indirectly, respectively [53,54]. According to the direct mode, cytochrome c interacts with apoptotic protease activating factor-1 (APAF1), thus transforming it into an apoptosome complex, which activates pro-caspase-9 to initiator caspase 9, which in turn activates caspase-3, leading to the activation of the final cascade and, consequently, the fragmentation of the nucleus after its membrane rupture [55]. After the activation of all signaling cascade mediators, the two pathways meet up at the final caspases’ activation (caspases 3, 7), resulting in the cleavage of different integrated cell proteins and the execution of the apoptotic steps [55].

Furthermore, the BCL-2 family of proteins [56], which tightly regulate the intrinsic pathway, is subclassified into three subgroups; the first comprises the pro-apoptotic proteins or apoptosis effectors that positively regulate apoptosis, and they include Bcl-2 antagonist killer 1 (Bak) and Bcl-2-associated X protein (Bax), which can directly execute MOMP when activated.

The second comprises the pro-survival members that negatively regulate apoptosis, and they include B cell lymphoma-2 (Bcl-2) itself, B cell lymphoma extra-large (Bcl-xL), B cell lymphoma-w (Bcl-W), B cell lymphoma-like protein 2 (Bcl-B), myeloid leukemia cell 1 (MCL-1) and Bcl-2-related protein A1/Bcl-2-related isolated from fetal liver-11 (A1/BFL-1).

The third group is the pro-apoptotic BH3-only proteins, which include p53-upregulated modulator of apoptosis (Puma), Bcl-2-interacting mediator of cell death (Bim) and the Bcl-2 Homology 3 interacting death agonist (Bid), which act as activators to Bax and Bak. This group comprises also other proteins, called sensitizers, which do not associate with Bax and Bak, but antagonize the pro-survival members, including the Bcl-2 associated agonist of cell death (Bad), Bcl-2 interacting t killer 1 (Bik), Noxa, BMF and HRK.

In healthy cells, the pro- and anti-apoptotic proteins are held in a fine, delicate balance. Conversely, during cancer development, the malignant cells alter this balance, by overexpressing various anti-apoptotic proteins and/or abnormally reduce the expression levels of pro-apoptotic members, and thus they become unresponsive to stimuli that otherwise trigger apoptosis in sensitive cells, which leads to cellular proliferation and cancer progression, in addition to therapeutic resistance [57]. Therefore, inhibiting the anti-apoptotic [58,59] and activating pro-apoptotic members [60] of the Bcl-2 family have been suggested as plausible selective strategies for cancer therapy and to overcome drug resistance.

Taken together, the complexity of CRC and the manifold and heterogeneous association of this malignancy with OS, inflammation, Warburg effect and hyperglycemia highlight the need for the development of new pharmaceuticals capable of the simultaneous targeting of these risk factors as well as the apoptotic pathways (Figure 1).

Figure 1. Various risk factors and pathways implicated in cancer, which serve as targets for multi-potent agents.

The innovation of multi-targeted therapies represents a promising and prevailing anticancer drug discovery paradigm among medicinal chemists [61]. In line with this principle, quinazolinone scaffolds have proven to be easily accessible pharmacophores, possessing multipotent pharmacological activities both in vitro and in vivo [62]. They have been presented as core structures in inhibitors for; AA [63] I, AG [64] II, and COX-2 [65] III, as well as antioxidant agent [66] IV, and anti-cancer candidates (V, VI and VII), which produce their cytotoxic effects through different molecular mechanisms [63,67,68], as portrayed in Figure 2.

Figure 2. Selected examples of biologically active compounds containing quinazolinone cores.

In light of the above, in this study, we designed a number of new quinazolinone derivatives bearing 4-nitro-phenyl, in addition to fluorine containing 4- or 3-(trifluoromethyl)phenyl moiety at position 3, due to the significant impact of fluorine atoms in enhancing the binding affinity to the targeted protein, as well as improving pharmacokinetic and physicochemical properties including metabolic stability and membrane permeation [69]. Also, various substituents such as Br, Cl and OCH₃ were located at the 6 and/or 6,7 positions on the quinazolinone core, to span different electronic, steric and lipophilic characters that would produce different pharmacological responses, which would increase the probability of the discovery of active agents. Subsequently, these compounds were evaluated in vitro as antioxidants as well as AA, AG, COX-2 and LDHA inhibitors. Structure–activity relationships regarding the antioxidant activity were discussed. Additionally, molecular docking simulations were performed to confirm the results of the in vitro enzymatic inhibitory assays and to predict the conformations of the potential inhibitors upon binding to the target proteins, as well as to calculate the binding affinities between them.

Moreover, all compounds were screened for their cytotoxic effects on LoVo and HCT-116 human colon carcinoma cells at 200 µg/mL, and then the IC₅₀ values of the most active cytotoxic candidates were calculated form the concentration–response curves and compared with that of 5-FU. Afterwards, they were evaluated against non-tumoral human umbilical vein endothelial cells (HUVEC) to explore their safety profiles. Furthermore, the capability of the cytotoxic candidates to modulate apoptosis regulators (Bax and BcL2), to activate caspases-8, 9 and 3 and to disturb the cell cycles of LoVo and HCT-116 cells were investigated. Finally, the physicochemical, medicinal chemistry and ADMT properties of all compounds were predicted and discussed in detail for some active compounds as model examples.

2. Results and Discussion

2.1. Chemistry

The target compounds were synthesized as depicted in Scheme 1.

Initially, refluxing mixtures of 5-substituted or 4,5-disubstituted anthranilic acid derivatives 1a-c and 3-substituted or /4-substituted phenyl isothiocyanate derivatives 2a-c in glacial acetic for 10 h [70] yielded the new 6-substituted or 6,7-disubstituted-3-aryl-2-thioxo-2,3-dihydro-1H-quinazolin-4-one derivatives 3a-h, which can potentially also exist in their tautomeric forms 3′a-h due to thiocarbonyl–thiol tautomerization. Subsequently, the arylation of the acidic thiol groups using various aryl halides 4a–d under basic catalysis with potassium carbonate in refluxing dry acetone for 6 h [50] produced eight new 2-(benzylsulfanyl)-3-aryl-3H-quinazolin-4-one derivatives, 5a–h. The structures of the synthesized derivatives were elucidated based on IR, ¹H-NMR, ¹³C-NMR and mass spectrometric analyses.

All the spectroscopic data indicated the predominance of the thiocarbonyl tautomeric form for compounds 3a–h in the solid state (IR) and in the DMSO solution (NMR), which is in agreement with the reported data [71]. Thus, their IR spectra showed stretching absorption bands for the NH groups at a ν_max ranging from 3155 to 3245 cm⁻¹. In addition, the ¹HNMR spectra demonstrated the NH protons as singlet peaks at δ_H = 12.91–13.24 ppm, while the ¹³CNMR spectra exhibited characteristic signals for C= S at a δ_C/ppm ranging from 174.38 to 176.28.

Contrarily, the IR and ¹HNMR spectra of derivatives 5a–h indicted the disappearance of the NH protons, and their ¹³CNMR spectra confirmed the absence of the peaks corresponding to the C=S groups and the presence of the benzylic methylene groups.

As representative examples, the IR spectrum of compound 3c showed the stretching absorption bands at ν_max = 3178 and 1698 cm⁻¹, corresponding to the NH and C=O groups, respectively. The ¹H-NMR spectrum of this compound (500 MHz; DMSO-d₆) showed a characteristic one-proton singlet peak at δ_H = 12.91 ppm for the NH proton. Additionally, the four aromatic protons of the 3-trifluoromethylphenyl ring exhibited a one-proton doublet peak (coupling constant J value = 7.5 Hz), a one-proton singlet peak, an apparent triplet peak (J value = 8.05 Hz) and a doublet peak ((J value = 8.5 Hz) at chemical shift values of δ_H = 7.73, 7.69, 7.66, 7.24 and 7.56 ppm, respectively. The two aromatic protons of the dimethoxyphenyl ring were detected as two singlet peaks at δ = 7.24 and 6.95 ppm.

Lastly, the two methoxy groups exhibited two singlet peaks, each integrating to three protons at δ_H = 3.83 and 3.77 ppm. With regard to its ¹³C-NMR spectrum (125 MHz; DMSO-d₆), it was characterized by a distinctive peak at δ_C/ppm = 174.66 attributed to C=S group, in addition to seventeen peaks, as expected, at δ_C/ppm = 159.34 (C=O), 155.41, 146.58, 140.23, 135.63, 133.62, 130.03, 129.80, 126.38, 125.03, 124.89, 122.87, 108.66, 106.94, 97.98 (6 × CH-aromatic, 6 × C_q-aromatic and C_qF₃), 56.01 and 55.84 (2 × OCH₃). Finally, the MS spectrum of this compound showed a molecular ion peak [M⁺ + 1] at m/z = 383.17 (27.26%) for C₁₇H₁₃F₃N₂O₃S, and the base peak was detected at m/z = 382.16, which corresponds to [M⁺].

On the other hand, the IR spectrum (KBr) of compound 5c indicated the disappearance of the stretching absorption band due to NH group of its precursor 3c. The ¹H-NMR spectrum (300 MHz, DMSO-d₆) endorsed the absence of NH proton and the presence of the characteristic benzylic protons as an AB quartet signal at δ_H = 4.53 ppm, with a coupling constant of J = 13.5 Hz. In addition, it displayed eight peaks corresponding to ten aromatic protons as follows: a one-proton apparent doublet of the doublet peak (J values = 6.3, 2.1 Hz); an apparent one-proton doublet of the doublet peak (J values = 8.4, 2.4 Hz); a one-proton singlet; a two-proton doublet peak (J = 7.8 Hz); a two-proton multiplet peak; a one-proton triplet peak (J = 7.8 Hz); a one-proton singlet peak and a one-proton singlet peak, which were resonating at δ_H = 8.52, 8.09, 7.98, 7.93, 7.83–7.77, 7.60, 7.37 and 7.24 ppm, respectively, confirming the installation of the 3-nitrophenyl moiety on the sulfur atoms. The ¹³C-NMR spectrum (150 MHz; DMSO-d₆) revealed the absence of the peak due to the C=S group and the presence a new characteristic CH₂-benzylic peak at δ_C = 35.12 ppm. The mass spectrum of this compound indicated the presence of the molecular ion peaks [M⁺ + 2] and [M⁺ + 1] at m/z (%) = 519.10 (6.53) and 518.19 (29.95), respectively, for C₂₄H₁₈F₃N₃O₅S. The base peak, which represents [M⁺] ion was recorded at m/z = 517.18.

2.2. Biological Evaluation

The synthesized compounds were subjected to multiple in vitro biological evaluations as described in the following sections.

2.3. Molecular Docking Simulations

In order to confirm the results concluded from the in vitro enzymatic inhibitory assays, molecular docking studies were performed. Thus, the relative binding affinities, binding interactions and conformations of the most active candidates within the active sites of their target proteins were identified.

2.4. In Silico Physicochemical, Medicinal and ADMET Predictions

The data for all the synthesized compounds are available in the .

3. Materials and Methods

3.1. Chemistry

3.2. Biology

3.3. In Silico Studies

4. Conclusions

Quinazolinone derivatives 3a–h and their S-arylated analogues 5a–h were synthesized and characterized by various spectroscopic techniques. Multi-target biological screening revealed the significance of 3a, 3g and 5a as potent DPPH radical scavengers with lowered IC₅₀ values (mM) of 0.191 ± 0.011, 0.165 ± 0.005 and 0.172 ± 0.004, respectively, as compared with 0.245 ± 0.027 by BHT. Despite this, none of the screened compounds showed an improved inhibitory efficiency to celecoxib (IC₅₀ = 0.136 × 10 ⁻³ ± 0.006 × 10 ⁻³ µM) and sodium oxamate (IC₅₀ = 140.503 ± 7.647 µM) against COX- 2 and LDHA, respectively; quinazolinones 3a (IC₅₀ values = 281.374 ± 10.545 and 273.345 ± 16.087 µM) and 3g (IC₅₀ values = 251.780 ± 22.023 and 242.279 ± 31.298) were recognized as the most active derivatives against these enzymes, respectively. Notably, quinazolinones 3h, 5a and 5h with IC₅₀ values ranging from 12.548 ± 0.542 to 12.882 ± 0.426 µM against AG, and derivatives 3a, 3c, 3f, 3h and 5a–5f as well as 5h, with IC₅₀ values spanning from 186.437 ± 9.700 to 381.335 ± 8.713 µM against AA, exerted superior activity to quercetin (IC₅₀ = 13.126 ± 0.688 and 402.566 ± 10.108, respectively). Therefore, 3h, 5a and 5h can be considered as dual inhibitors for AG and AA.

In addition, the viability assays identified 3a and 3f as the most active antiproliferative agents with IC₅₀ (µM) values of 294.324 ± 8.409 and 383.521 ± 8.989 (LoVo cells), and 298.060 ± 13.247 and 323.596 ± 2.996(HCT-116 cells) as compared with 22.371 ± 0.215(LoVo) and 91.098 ± 2.721(HCT-116) by 5-FU. Interestingly, these compounds did not inhibit the growth of non-tumoral HUVEC at 560.617 and 599.251 µM, with viability percents of 97.000 ± 2.646 and 99.667 ± 0.577, respectively. Quantitative real-time PCR measurements confirmed the capability of 3a and 3f at 10 µg/mL to upregulate the expression levels of Bax and Caspase-3 genes, along with the downregulation of the expression level of the Bcl-2 gene in the cultures of the treated HCT-116 and LoVo cells as compared to the negative control cultures. Moreover, the Western blotting analyses indicated that 3a and 3f were able to induced apoptosis in HCT-116 and LoVo colon cancer cells via pathways triggered by caspases, and predominantly through the intrinsic pathway mediated by caspase-9. Furthermore, the two compounds induced more death for the HCT-116 and LoVo cells via apoptosis than via necrosis; in addition, they induced cell cycle arrest for the LoVo cells in the G2/M phase and for the HCT-116 cells in the G1 phase. Therefore, the observed cytotoxicity of these derivatives could be attributed to the induction of apoptosis and the blocking of cell cycle progression in both cell lines. Molecular docking analyses of the active or potential candidates corroborated the observed inhibitory efficiencies in the in vitro assays against AA, AG, LDAH and COX-2. Lastly, the physicochemical properties, the suitability in medicinal chemistry and the ADMET characteristics were predicted for all the synthesized compounds and explained in detail for some representative active compounds. Collectively, these quinazolinone derivatives may be considered to be suitable candidates for further optimization and exploration against CRC and its risk factors, particularly oxidative stress and DM-type 2.

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