Bio-Oriented Synthesis and Molecular Docking Studies of 1,2,4-Triazole Based Derivatives as Potential Anti-Cancer Agents against HepG2 Cell Line

Triazole-based acetamides serve as important scaffolds for various pharmacologically active drugs. In the present work, structural hybrids of 1,2,4-triazole and acetamides were furnished by chemically modifying 2-(4-isobutylphenyl) propanoic acid (1). Target compounds 7a–f were produced in considerable yields (70–76%) by coupling the triazole of compound 1 with different electrophiles under different reaction conditions. These triazole-coupled acetamide derivatives were verified by physiochemical and spectroscopic (HRMS, FTIR, 13CNMR, and 1HNMR,) methods. The anti-liver carcinoma effects of all of the derivatives against a HepG2 cell line were investigated. Compound 7f, with two methyl moieties at the ortho-position, exhibited the highest anti-proliferative activity among all of the compounds with an IC50 value of 16.782 µg/mL. 7f, the most effective anti-cancer molecule, also had a very low toxicity of 1.190.02%. Molecular docking demonstrates that all of the compounds, especially 7f, have exhibited excellent binding affinities of −176.749 kcal/mol and −170.066 kcal/mol to c-kit tyrosine kinase and protein kinase B, respectively. Compound 7f is recognized as the most suitable drug pharmacophore for the treatment of hepatocellular carcinoma.


Introduction
In the 21st century, cancer and other infectious diseases are the most prevalent causes of death globally [1]. According to the World Health Organization (WHO), 11.5 million deaths are expected by 2030 due to cancer [2]. Among all types of cancer, hepatocellular carcinoma is among the leading causes of death, accounting for approximately 92% mortality rates

Introduction
In the 21st century, cancer and other infectious diseases are the most prevalent causes of death globally [1]. According to the World Health Organization (WHO), 11.5 million deaths are expected by 2030 due to cancer [2]. Among all types of cancer, hepatocellular carcinoma is among the leading causes of death, accounting for approximately 92% mortality rates worldwide [3,4]. Thus, the development of new anti-cancer drugs remains a huge clinical need for improving therapeutic efficacy and controlling cancer [5]. The development of multi-target anti-cancer agents is the major focus of researchers globally due to the different drawbacks associated with already-used chemotherapeutics such as undesirable side effects, a lack of selectivity, systemic toxicity, and the emergence of multidrug resistance [6][7][8][9]. Intensive efforts must be made to discover and develop new, effective, tailored -anti-cancer agents with better safety profiles and drug-like properties.
Structurally modified nitrogen-containing heterocyclic moieties have a broad spectrum of applications for the development of novel therapeutic drugs as shown in Figure  1 [10,11]. Approximately 75% of Food and Drug Administration-approved drugs are nitrogen-based moieties [12]. Nitrogen-containing heterocyclic compounds have been synthesised in large numbers in recent times. They exhibit anti-tubercular [13], anti-cancer [14], anti-fungal [15], anti-microbial [16], anti-viral [17], and other biological properties such as genotoxicity and lipid peroxidation [18], and anti-inflammatory properties [19]. Molecular hybridization is an easy and efficient method to combine various important drug pharmacophores. Our ongoing research focuses on the design and synthesis of pharmacologically active, diverse polyvalent scaffolds as anti-cancer agents. The versatile nature of 1,2,4-triazole has been reported to be of great importance in medicinal chemistry, such as for its anti-cancer [20], anti-fungal [21], anti-bacterial [22], anti-microbial, and anti-tumor properties [23], as well as pyrophosphatases and phosphodiesterase [24]. Acetamide has been identified as the most significant pharmacophore of anti-cancer drugs [25].
On this basis, we have created a hybrid of acetamide and 1,2,4-triazole pharmacophore by chemical derivatization of 2-(4-isobutylphenyl)propanoic acid in an attempt to avoid tumor progression. On each side of triazole and acetamide, we developed a Molecular hybridization is an easy and efficient method to combine various important drug pharmacophores. Our ongoing research focuses on the design and synthesis of pharmacologically active, diverse polyvalent scaffolds as anti-cancer agents. The versatile nature of 1,2,4-triazole has been reported to be of great importance in medicinal chemistry, such as for its anti-cancer [20], anti-fungal [21], anti-bacterial [22], anti-microbial, and antitumor properties [23], as well as pyrophosphatases and phosphodiesterase [24]. Acetamide has been identified as the most significant pharmacophore of anti-cancer drugs [25].
On this basis, we have created a hybrid of acetamide and 1,2,4-triazole pharmacophore by chemical derivatization of 2-(4-isobutylphenyl)propanoic acid in an attempt to avoid tumor progression. On each side of triazole and acetamide, we developed a molecular framework with hydrophobic aryl rings. It improves the solubility of the drug and its candidacy as a good drug molecule. Earlier, we reported the synthesis of various structural hybrids of oxadiazole-based acetamide [26][27][28][29], and it has been proven from the literature that heterocycle-based compounds possess good anti-cancer activity [30]. Thus, in an extension of our earlier research on heterocycles, -COOH group of 2-(4-isobutylphenyl)propanoic acid was cyclized into a 1,2,4-triazole ring 4.

Chemistry
In the current study, 2-(4-isobutylphenyl)propanoic acid has been chemically modified with improved clinical utility. Different N-arylated 5-aryl-1,2,4-triazole-coupled acetamides (7a-f) have been synthesized in good yields by replacement of the H group of SH with various electrophiles.

Anti-proliferative Potential
The anti-hepatocellular activity of afforded N-arylated 1,2,4-triazole coupled acetamides (7a-f) was evaluated via MTT assay and these structural hybrids were screened against a liver cancer HepG2 cell line [33]. All of the compounds demonstrated mild to Scheme 1. Synthesis of 1,2,4-triazole-cored acetamides 7a-f. In the 1 HNMR spectrum, -NH protons of the acetamide were the most deshielded and their chemical shift value was observed around 10.27-9.64 ppm. In propanoic acid, the protons of the aliphatic region were among the most shielded, with values ranging from 0.86-0.84 ppm. The presence of acetamide was confirmed by the appearance of signals around 4.04 ppm for the CH 2 group and in the range of 10.27-9.64 ppm for the NH group. In 13 CNMR, the appearance of C=O signals in the range of 164.15-168 ppm confirmed the synthesis of acetamide. The presence of a triazole ring in the final derivatives was also confirmed by the appearance of signals in the range of 158.15 ppm and at 29.58 ppm for N-CH 3 . In the 1 HNMR spectrum, signals for N-CH 3 were observed around 3.28 ppm. A distinctive peak around 22.0 ppm corresponded to the protons of two -CH 3 carbon nuclei. Signals for the CH 2 groups were observed between 44.18 to 20.19 ppm. By introducing some electron-withdrawing substitutions and comparing them to electron-donating group substitutions, we reported structure-activity relationships for the phenyl group. 3,5-disubstituted triazole nuclei have a versatile nature and are important in the pharmaceutical industry. On the basis of their medicinal importance, the anti-cancer activities of all of the compounds were checked.

Anti-proliferative Potential
The anti-hepatocellular activity of afforded N-arylated 1,2,4-triazole coupled acetamides (7a-f) was evaluated via MTT assay and these structural hybrids were screened against a liver cancer HepG2 cell line [33]. All of the compounds demonstrated mild to outstanding anti-cancer activity, as shown in Table 1. Among all of the compounds, 7f, with two methyl groups at positions 2 and 6 of the phenyl ring, displayed the best anti-cancer potential with IC 50 = 16.782 µg/mL. Compound 7a, which contains a methyl at position 2 of the phenyl ring, also displayed good anti-cancer activity with an IC 50 value of 20.667 µg/mL but less than 7f. Compounds 7b, 7c, and 7e also exhibited a significant anti-cancer effect but less than 7f and 7a. Compound 7d, with an electron-withdrawing Cl substituent, displayed the lowest anti-hepatocellular activity with a 39.667 µg/mL IC 50 value. The cell viability of all of the compounds was further evaluated using various concentrations (3.125-200 µg) to test the dose response and % inhibition relationship as shown in Table 2. Figure 2 shows that compounds 7f and 7a produced the best results at a 25 µg/mL dose among all of the compounds.
The dose response and % inhibition of the most potent compound, 7f, was checked at various concentrations ( Figure 3).  The dose response and % inhibition of the most potent compound, 7f, was checked at various concentrations ( Figure 3).

Structure-Activity Relationship of 7a-f
The anti-hepatocellular potential of all of the derivatives, 7a-f, was evaluated against a HepG2 cancer cell line at various concentrations via MTT assay. The most potent derivative was 7f, which contains two methyl groups at ortho-positions of the phenyl ring (IC50 value of 16.782 µM). Compound 7d, which contained an electron-withdrawing Cl-group in an orthogonal position, showed the lowest anti-cancer potency with an IC50 value of 39.667 µ M. The anti-proliferative activity of all of the derivatives was decreased in the following order: 7f > 7a > 7b > 7c > 7e > 7d. This proves that the attachment of an electrondonating CH3 group at the ortho-position increases the anti-cancer activity of the compounds. Based on the results of the SAR of N-arylated 5-aryl-1,2,4-triazole-coupled acetamide scaffolds 7a-f, it was determined that the -CH3 motif at the ortho-position of the phenyl ring improved the anti-proliferative potential of the compounds.

Molecular Docking
Molecular docking screenings were carried out to theoretically predict the most promising protein targets of compounds by molecular docking to some cancer targets. Five major targets in the treatment of cancer have been identified: human Aurora B kinase, phosphatidylinositol 3-kinase alpha (PI3Kalpha), the signal transducer and activator of transcription 3 (STAT3), protein kinase B (Akt), and c-kit tyrosine kinase (c-Kit). The web page http://www.swisstargetprediction.ch/ (Accessed on 7 November 2022) was used to

Structure-Activity Relationship of 7a-f
The anti-hepatocellular potential of all of the derivatives, 7a-f, was evaluated against a HepG2 cancer cell line at various concentrations via MTT assay. The most potent derivative was 7f, which contains two methyl groups at ortho-positions of the phenyl ring (IC 50 value of 16.782 µM). Compound 7d, which contained an electron-withdrawing Cl-group in an orthogonal position, showed the lowest anti-cancer potency with an IC 50 value of 39.667 µM. The anti-proliferative activity of all of the derivatives was decreased in the following order: 7f > 7a > 7b > 7c > 7e > 7d. This proves that the attachment of an electron-donating CH 3 group at the ortho-position increases the anti-cancer activity of the compounds. Based on the results of the SAR of N-arylated 5-aryl-1,2,4-triazole-coupled acetamide scaffolds 7a-f, it was determined that the -CH 3 motif at the ortho-position of the phenyl ring improved the anti-proliferative potential of the compounds.

Molecular Docking
Molecular docking screenings were carried out to theoretically predict the most promising protein targets of compounds by molecular docking to some cancer targets. Five major targets in the treatment of cancer have been identified: human Aurora B kinase, phosphatidylinositol 3-kinase alpha (PI3Kalpha), the signal transducer and activator of transcription 3 (STAT3), protein kinase B (Akt), and c-kit tyrosine kinase (c-Kit). The web page http://www.swisstargetprediction.ch/ (Accessed on 7 November 2022) was used to study the potential anti-cancer effect of new synthesized molecules [35]. The results suggest that molecules may be effective against kinase targets. Some important kinases and important targets in cancer treatment were determined by the literature review. c-kit tyrosine kinase endorses cellular proliferation by activating signal transduction mechanisms in response to stem cell factor adhesion [36]. Akt plays a vital role in internal cell signalling by accelerating cellular survival and proliferation. Its path becomes irregular during cancer [37]. Aurora kinase B regulates the cell cycle and is ubiquitously expressed in cancerous cells. [38]. The insulin-like growth factor-1 receptor (IGF-1R) has a vital role in cells in conjunction with PI3K-AKT and Ras-Raf-MEK signalling cascades, which control proliferation and apoptosis within cells. It is considered an important therapeutic target because of its deregulation of solid tumor types [39]. Phosphatidylinositol 3-kinase alpha is an intracellular lipid kinase that regulates cell survival, development, proliferation, and metabolism. It has been linked to a number of human cancers [40]. STAT3 is a secret transcription factor; it is regarded as an appealing target of anti-cancer therapeutics [41]. Table 3 shows the selected targets, grid box coordinates, their protein data bank codes, and Moldock scores. The docking findings demonstrate that molecules have the ability to affect a variety of targets. c-kit and Akt specifically are anticipated to have a high binding potential with cancer therapeutic targets, as demonstrated in Table 3.   The docking findings demonstrate that all of the compounds have good binding affinity to kinase protein. The protein c-kit tyrosine kinase is expected to interact strongly with cancer-targeted therapeutics. Table 4 shows the docked complexes' categories, modes of interactions, binding affinities, and by-products. Each ligand's hydrophobic  The docking findings demonstrate that all of the compounds have good binding affinity to kinase protein. The protein c-kit tyrosine kinase is expected to interact strongly with cancer-targeted therapeutics. Table 4 shows the docked complexes' categories, modes of interactions, binding affinities, and by-products. Each ligand's hydrophobic contacts and hydrogen bonding interactions were evaluated within the receptor protein's binding pocket. Compound 7f bonded to c-kit tyrosine kinase with the most suitable binding pose and a low binding energy of −176.749 kcal/mol. It formed an H-bond with Asp810 and Glu640. Hydrophobic interactions occurred such as the pi-sigma bond with Thr670, His790, and Val643. Other hydrophobic interactions involved alkyl interactions with Val603, Val668, Leu644, Leu783, and Ile571, pi-sulfur interactions with Cys788 and Lys623, and amidepi-stacked interactions with Cys809 residue. It also interacts with Val654, Ile789, Leu647, Ile808, Ile653, and Phe811 residues through van der Waals interaction. The 2D and 3D diagrammatic expressions of the binding interactions of 7f and c-kit tyrosine kinase are shown in Figure 5.
Glu640. Hydrophobic interactions occurred such as the pi-sigma bond with Thr670, His790, and Val643. Other hydrophobic interactions involved alkyl interactions with Val603, Val668, Leu644, Leu783, and Ile571, pi-sulfur interactions with Cys788 and Lys623, and amide-pi-stacked interactions with Cys809 residue. It also interacts with Val654, Ile789, Leu647, Ile808, Ile653, and Phe811 residues through van der Waals interaction. The 2D and 3D diagrammatic expressions of the binding interactions of 7f and ckit tyrosine kinase are shown in Figure 5. Compound 7a combines with c-kit tyrosine kinase with the most suitable binding poses with a low binding energy of −173.411 kcal/mol. Hydrophobic interactions of 7a involved a pi-sigma bond with Leu595 and Leu644 and alkyl interactions with Cys673, Cys809, Ala621, Val654, Tyr672, and Phe811. It also interacts with Gly676, Leu799, Asp810, Leu647, Leu783, His790, Ile808, Glu640, Thr670, Val603, and Asp677 residues by van der Waals interactions. The 2D and 3D diagrammatic expressions of the binding interactions of 7a and c-kit tyrosine kinase are shown in Figure 6. The docked conformations of reference ligand X39 and protein kinase B were investigated in order to calculate qualitative estimates and the molecular basis of the analyzed Compound 7a combines with c-kit tyrosine kinase with the most suitable binding poses with a low binding energy of −173.411 kcal/mol. Hydrophobic interactions of 7a involved a pi-sigma bond with Leu595 and Leu644 and alkyl interactions with Cys673, Cys809, Ala621, Val654, Tyr672, and Phe811. It also interacts with Gly676, Leu799, Asp810, Leu647, Leu783, His790, Ile808, Glu640, Thr670, Val603, and Asp677 residues by van der Waals interactions. The 2D and 3D diagrammatic expressions of the binding interactions of 7a and c-kit tyrosine kinase are shown in Figure 6.
Glu640. Hydrophobic interactions occurred such as the pi-sigma bond with Thr670, His790, and Val643. Other hydrophobic interactions involved alkyl interactions with Val603, Val668, Leu644, Leu783, and Ile571, pi-sulfur interactions with Cys788 and Lys623, and amide-pi-stacked interactions with Cys809 residue. It also interacts with Val654, Ile789, Leu647, Ile808, Ile653, and Phe811 residues through van der Waals interaction. The 2D and 3D diagrammatic expressions of the binding interactions of 7f and ckit tyrosine kinase are shown in Figure 5. Compound 7a combines with c-kit tyrosine kinase with the most suitable binding poses with a low binding energy of −173.411 kcal/mol. Hydrophobic interactions of 7a involved a pi-sigma bond with Leu595 and Leu644 and alkyl interactions with Cys673, Cys809, Ala621, Val654, Tyr672, and Phe811. It also interacts with Gly676, Leu799, Asp810, Leu647, Leu783, His790, Ile808, Glu640, Thr670, Val603, and Asp677 residues by van der Waals interactions. The 2D and 3D diagrammatic expressions of the binding interactions of 7a and c-kit tyrosine kinase are shown in Figure 6. The docked conformations of reference ligand X39 and protein kinase B were investigated in order to calculate qualitative estimates and the molecular basis of the analyzed The docked conformations of reference ligand X39 and protein kinase B were investigated in order to calculate qualitative estimates and the molecular basis of the analyzed bioactive molecules. Figure 7 shows 2D and 3D diagrammatic representations of the molecular bindings between the reference ligand X39 and kinase B protein's active pocket. bioactive molecules. Figure 7 shows 2D and 3D diagrammatic representations of the molecular bindings between the reference ligand X39 and kinase B protein's active pocket. According to the docking findings, all triazole-coupled acetamide scaffolds have a significant ability to influence the protein kinase B moiety. Akt is anticipated to have a high affinity for carcinoma therapeutic targets. Table 3 shows the docked complexes' binding interactions, classifications, kinds of interactions, and interacting residues. Each According to the docking findings, all triazole-coupled acetamide scaffolds have a significant ability to influence the protein kinase B moiety. Akt is anticipated to have a high affinity for carcinoma therapeutic targets. Table 3 shows the docked complexes' binding interactions, classifications, kinds of interactions, and interacting residues. Each ligand's hydrophobic contacts and hydrogen bonding interactions were assessed within the receptor protein's binding site. Table 5 describes the ligand conformations that demonstrated the greatest biological activity, as well as their suitable interactions in the receptors. Compound 7a had the most suitable binding poses with a binding energy of −166.843 kcal/mol to protein kinase B. Compound 7a had the most suitable binding poses with a low energy of −166.843 kcal/mol to the catalytic site of protein kinase B. 7a bonded to protein kinase B via H-bonding with Gly164, Phe163, and Thr162, a carbon-hydrogen bond with SerC9, pi-sigma interaction with Gly161, and pi-sulfer interaction with Asp293. It bonded with residues Leu183, val166, and Lys181 via hydrophobic alkyl interactions. Asn280, Lys277, Glu279, ArgC6, Lys160, Glu236, Gly159, Lys165, and ThrC8 residues involve van der Waals interactions among protein kinase B and compound 7a (Figure 8).

General
In the present research, all of the starting materials were of analytical grade and were purchased from Alfa Aesar or Sigma Aldrich. 2-(4-isobutylphenyl) propanoic acid was used as a starting material. A Stuart SMP10 melting point apparatus was used for determining the melting point of all of the derivatives. The structures of all of the triazole-based scaffolds were confirmed by spectroscopy and physiochemical methods. An FT-IR spectrophotometer (4000-400 cm −1 ) by BRUKER was used at the Hi-Tech Lab, GC University, Faisalabad. 1 HNMR spectra were recorded on an Bruker Advance 500 MHz spectrophotometer using DMSO-d6, on 5 mm diameter tubes at the University of Copenhagen, Denmark. An Bruker Advance NMR spectrophotometer was used to record 13 C NMR spectra at 75 MHz by using DMSO-d6, on 5 mm diameter tubes. The reactions were supervised by thin layer chromatography.

General
In the present research, all of the starting materials were of analytical grade and were purchased from Alfa Aesar or Sigma Aldrich. 2-(4-isobutylphenyl) propanoic acid was used as a starting material. A Stuart SMP10 melting point apparatus was used for determining the melting point of all of the derivatives. The structures of all of the triazole-based scaffolds were confirmed by spectroscopy and physiochemical methods. An FT-IR spectrophotometer (4000-400 cm −1 ) by BRUKER was used at the Hi-Tech Lab, GC University, Faisalabad. 1 HNMR spectra were recorded on an Bruker Advance 500 MHz spectrophotometer using DMSO-d6, on 5 mm diameter tubes at the University of Copenhagen, Denmark. An Bruker Advance NMR spectrophotometer was used to record 13 C NMR spectra at 75 MHz by using DMSO-d6, on 5 mm diameter tubes. The reactions were supervised by thin layer chromatography.  (4) In the current study, methyl isothiocyanate and 2-(4-isobutylphenyl) propane hydrazide (0.02 mol) were dissolved in 10% KOH soln. in an equimolar amount. For 10-11 h, the mixture was set on refluxing at 95 • C. Thin-layer chromatography was used for monitoring the reaction. Upon completion, cold water was added to afford the precipitates of product. Water was used to filter and wash the precipitates. The precipitates were further purified with an ethanolic recrystallization process.

Synthesis of N-Aryl/Alkyl 2-Bromoroacetamides 6a-f
Compounds 6a-f were synthesized using the reported method [27]. In an RBF, 12.0 moles N-substituted alkyl/aryl amines (5a-f) were dissolved in 10.0 mL of 5% Na 2 CO 3 solution. Bromoacetyl bromide (12.0 mmoles) was gradually added to the reaction mixture described above. Upon reaction completion, n-hexane was added to afford arylated derivatives as precipitates which were further purified with an ethanolic recrystallization process or column chromatography technique using ethyl acetate-petroleum ether (1:9).
Human HepG2 liver cancer cell lines were cultured by Dulbecco's modified Eagle's medium. It is composed of 100 µg/mL streptomycin, 100 units/mL penicillin, and 10% FBS. A humidified atmosphere was provided for incubation at 37 • C with 5% CO 2 . The antihepatocellular therapeutic potential of triazole-based scaffolds was evaluated by dissolving its different concentrations in 0.05% DMSO.

Evaluation of Cell Viability
An MTT assay was applied for evaluation of cell viability against the HepG2 cell line [44]. In short, different concentrations of new triazole-based scaffolds were incubated with HepG2 cell lines for 48 h. After incubation, 5 mg/mL of 10 µL MTT solution was added in each plate and they were further incubated at 37 • C for 4 h. The percentage of cell viability was calculated at 490 nm after the addition of 150 µL DMSO into a microplate reader (Thermo Scientific, Waltham, MA, USA).

Hemolytic Activity Potential
Hemolytic activity was investigated by the reported method [45,46] using Triton-X-100 as standard.

Molecular Docking of Triazole-Coupled Acetamides
Docking experiments for all of the scaffolds were carried out in order to comprehend the potential interaction process of the synthesized anti-cancer compounds on the HepG2 cancer cell line. The website https://www.rcsb.org was used for drawing the structures of PI3Kalpha, Akt, c-kit tyrosine kinase, human Aurora B kinase, and STAT3 from the RCSB Protein Data Bank under the PDB IDs of 4FA6, 2X39, 1T46, 4AF3, and 6NJS, respectively [36][37][38][39]47]. ChemDraw 20.1.1 was used to create and reduce the 3D SDF structures of all of the compounds, which were then transferred to MarvinSketch. Prior to docking, the target proteins' frameworks were evaluated, and errors in amino acid structures were rectified using Molegro Virtual Docker software [48]. The grid boxs' centers were chosen to be the co-crystallized ligands of proteins. They re-docked in order to validate the in silico process. Molegro Virtual Docker was applied to dock active chemicals 10 times to the target proteins' receptors. The sequences with the lowest interaction affinity and excellent connections with the targets were separated for further detailed analysis. The molecular bindings between the target and new derivatives were visualized in 2D using Discovery Studio Visualizer Software 2021.

Conclusions
A series of new anti-cancer compounds (7a-f) were synthesized in moderate to good yield (73-76%) by combining compound 4 with various electrophiles under different reaction conditions (Table 1, Scheme 1). Because of its low bioavailability of 38-49%, Sorafenib necessitates a significant daily dose in cancer therapy. Sorafenib is a very costly medicine with many side effects. We have incorporated various electron-donating and electron-withdrawing groups into electrophiles to test structure-activity relationships at various concentrations. All of the molecules demonstrated medium to outstanding anti-cancer activity, comparable to sorafenib, which diversified according to aryl ring substitution, as shown in Table 1. These triazole-based acetamide derivatives also exhibited low cytotoxicity, with values ranging from 7.33% to 1.19% in comparison to the 100% cytotoxicity exhibited by the reference standard Triton X100. Compounds 7f and 7a showed the highest anti-cancer potential, with IC 50 values of 16.782 µg/mL and 20.667 µg/mL, respectively. On the other hand, the triazole derivative containing an electron-withdrawing chloro moiety demonstrated the least anti-proliferative activity with an IC 50 value of 39.667 µg/mL. The sequence of anti-cancer potential was found to be 7f > 7a > 7b > 7c > 7e > 7d. The anti-cancer potential of all of the compounds was further investigated by molecular docking studies and the results were in accordance with in-vitro studies. In silico studies have shown that the molecules have strong affinity for kinase targets. Molecules 7f and 7a have shown their anti-cancer effects, especially by affecting Akt and c-lit molecular targets. According to in silico modelling studies, 7f has an outstanding docking score with the lowest binding energy of −170.066 kcal/mol, which is lower than the reference ligand X39 for protein kinase B (−130.624 kcal/mol). We concluded that compound 7f contained electron-donating methyl groups at the 2 and 6 position of the aryl ring and showed good anti-cancer activity, low cytotoxicity, and good thrombolytic activity. Thus, compound 7f might be utilized to synthesize new anti-cancer drugs in the near future.