Biological Screening Research Articles

Rocephin-graphene oxide-silver nanocomposites: A versatile platform for biomedical applications.

For many years, the synthesis of graphene oxide (GO) had involved exfoliating graphite flakes, and the methods applied were expensive and time-consuming. Thus, an attempt had been made to create an inventive, less expensive method for the synthesis of GO using unrefined, raw carbon-containing material. Modified Hummer's method was used to prepare GO from banana peel. In addition, the metallic silver nanocomposite was also synthesized along with laoding of drug Rocephin where they interact with each other through electrostatic hydrogen bond interaction. The degree of crystallinity and the crystallite size were through x-ray diffraction (XRD) analysis and the crystallite size of AgNPs was found to be 40.40 nm. The scanning electron microscopy (SEM) analysis shows that the morphology of the GO gradually changes with the addition of AgNPs and Rocephin. A blue shift was seen in the absorbance maxima of the raw carbon upon the conjugation of Rocephin in UV analysis. The Fourier-transform infrared spectroscopy, and energy dispersive X-ray (EDX) spectroscopy were used to determine the chemical composition of the samples. Furthermore, a broad biological screening of the synthesized samples had been carried out following the total reducing power (TRP), total antioxidant capacity (TAC), antibacterial, antifungal, MTT (Cytotoxicity of biologically synthesized silver nanoparticles in MDA-MB-231 human breast cancer cells) cell viability, brine shrimp lethality, and hemolytic protocols. Significant results were obtained, and the Rocephin-GO-AgNPs had depicted promising activity as compared with their counterparts. RESEARCH HIGHLIGHTS: The GO was prepared from the raw carbon extracted from banana peels and was used as a substrate for the synthesis Graphene oxide silver nanoparticles (GO-AgNPs) and Rocephin-loaded graphene oxide silver nanoparticles (Rocephin-GO-AgNPs) The structural and compositional analysis of the nanomaterial was carried out, and they were screened for several biomedical applications. The Rocephin-GO-AgNPs exhibit the highest activity as compared with their counterparts.

Biological screening to identify hits the Therapeutic Targets of Alzheimer's disease and their role in the pathogenesis

Alzheimer's disease (AD) is a major problem in today's societies. More than five million Americans are living with Alzheimer's disease in the United States, with the majority being 65 and older. According to the Alzheimer's Association Report, the number of persons affected by Alzheimer's disease in the United States would rise to fourteen million by 2060. The disease, which is the most prevalent form of dementia, is a progressive and irreversible brain disorder that gradually deteriorates an individual's cognitive function. It advances from preclinical to early- to moderate- to late-stage disease. Early symptoms primarily include cognitive impairment, particularly memory loss. Current Alzheimer's disease treatment can be divided into two categories based on the disease's stage. Galantamine, rivastigmine, and donepezil as acetylcholinesterase inhibitors are suitable for mild to moderate cases to provide transient symptomatic relief among patients. Memantine, an N-methyl D-aspartate (NMDA) antagonist, is used as monotherapy to treat symptoms of moderate to severe Alzheimer's disease. These medications are typically selective molecules that target certain proteins ("one compound-one target" method), and their main goal is to restore physiological acetylcholine levels. Nonetheless, multiple pathways of Alzheimer's disease pathogenesis have been hypothesized to far, and they have been proven to overlap and influence one another.

Comprehensive Data-Driven Assessment of Non-Kinase Targets of Inhibitors of the Human Kinome.

Protein kinases (PKs) are involved in many intracellular signal transduction pathways through phosphorylation cascades and have become intensely investigated pharmaceutical targets over the past two decades. Inhibition of PKs using small-molecular inhibitors is a premier strategy for the treatment of diseases in different therapeutic areas that are caused by uncontrolled PK-mediated phosphorylation and aberrant signaling. Most PK inhibitors (PKIs) are directed against the ATP cofactor binding site that is largely conserved across the human kinome comprising 518 wild-type PKs (and many mutant forms). Hence, these PKIs often have varying degrees of multi-PK activity (promiscuity) that is also influenced by factors such as single-site mutations in the cofactor binding region, compound binding kinetics, and residence times. The promiscuity of PKIs is often-but not always-critically important for therapeutic efficacy through polypharmacology. Various in vitro and in vivo studies have also indicated that PKIs have the potential of interacting with additional targets other than PKs, and different secondary cellular targets of individual PKIs have been identified on a case-by-case basis. Given the strong interest in PKs as drug targets, a wealth of PKIs from medicinal chemistry and their activity data from many assays and biological screens have become publicly available over the years. On the basis of these data, for the first time, we conducted a systematic search for non-PK targets of PKIs across the human kinome. Starting from a pool of more than 155,000 curated human PKIs, our large-scale analysis confirmed secondary targets from diverse protein classes for 447 PKIs on the basis of high-confidence activity data. These PKIs were active against 390 human PKs, covering all kinase groups of the kinome and 210 non-PK targets, which included other popular pharmaceutical targets as well as currently unclassified proteins. The target distribution and promiscuity of the 447 PKIs were determined, and different interaction profiles with PK and non-PK targets were identified. As a part of our study, the collection of PKIs with activity against non-PK targets and the associated information are made freely available.

An improved method for toxicological profiling of chemical substances

Toxicity profiling is an integral part of the drug discovery pipeline. The 3Rs principle—Replacement, Reduction, and Refinement, is considered a golden rule in determining the most appropriate approach for toxicity studies. The acute toxicity study with proper estimate of median lethal dose (LD50) is usually an initial procedure for the determination of most suitable test doses for preclinical toxicological and pharmacological profiling. Several methods, which have been devised to determine the LD50, are faced with the challenge of using a large number of animals and time constraints. Despite the inherent advantage of the newer OECD Test Guidelines, the increasing concerns among toxicologists, the regulatory authorities and the general public, on the need to adhere to 3Rs principle, necessitated the need for an improved approach. Such an approach should not only minimize the time and number of animals required, but also take into cognizance animal welfare, and give accurate, comparable, and reproducible results across laboratories. While taking advantage of the inherent merits of the existing methods, here is presented the mathematical basis and evaluation of an improved method for toxicity profiling of test substances and estimation of LD50. The method makes use of the generated Table of values for the selection of appropriate test doses. Our proposed method has capacities to optimize the time and number of animal use, ensure more reliable and reproducible results across laboratories, allow for easy selection of doses for subsequent toxicity profiling, and be adaptable to other biological screening beyond toxicity studies.

Novel 5-Substituted Oxindole Derivatives as Bruton's Tyrosine Kinase Inhibitors: Design, Synthesis, Docking, Molecular Dynamics Simulation, and Biological Evaluation.

Bruton's tyrosine kinase (BTK) is a non-RTK cytoplasmic kinase predominantly expressed by hemopoietic lineages, particularly B-cells. A new oxindole-based focused library was designed to identify potent compounds targeting the BTK protein as anticancer agents. This study used rational approaches like structure-based pharmacophore modeling, docking, and ADME properties to select compounds. Molecular dynamics simulations carried out at 20 ns supported the stability of compound 9g within the binding pocket. All the compounds were synthesized and subjected to biological screening on two BTK-expressing cancer cell lines, RAMOS and K562; six non-BTK cancer cell lines, A549, HCT116 (parental and p53-/-), U2OS, JURKAT, and CCRF-CEM; and two non-malignant fibroblast lines, BJ and MRC-5. This study resulted in the identification of four new compounds, 9b, 9f, 9g, and 9h, possessing free binding energies of -10.8, -11.1, -11.3, and -10.8 kcal/mol, respectively, and displaying selective cytotoxicity against BTK-high RAMOS cells. Further analysis demonstrated the antiproliferative activity of 9h in RAMOS cells through selective inhibition of pBTK (Tyr223) without affecting Lyn and Syk, upstream proteins in the BCR signaling pathway. In conclusion, we identified a promising oxindole derivative (9h) that shows specificity in modulating BTK signaling pathways.

Targeting the Antifungal Activity of Carbon Dots against Candida albicans Biofilm Formation by Tailoring Their Surface Functional Groups.

Carbon dots (CDs) are an emerging class of carbon nanoparticles, which for their characteristics have found applications in many fields such as catalysis, materials and biomedicine. Within this context, the application of CDs as antibacterial agents has received much attention in very recent years, while their use as antifungal nanoparticles has been scarcely investigated. Here we report a systematic investigation of the surface functional groups of CDs to study their influence on these nanoparticles' against Candida albicans. Three classes of CDs have been synthesised and fully characterized. A thorough in vitro and in vivo biological screening against C. albicans was performed to test their antifungal, antiadhesion and antibiofilm formation activities. Moreover, the interaction with C. albicans cells was investigated by microscopic analysis. Our results evidence how the presence of a positively polarised surface results crucial for the internalization into COS-7 cells. Positively charged nanoparticles were also able to inhibit adhesion and biofilm formation, to interact with the cellular membrane of C. albicans, and to increase the survival of G. mellonella infected larvae after the injection with positive nanoparticles. The antifungal activity of CDs and their extremely low toxicity may represent a new strategy to combat infections sustained by C.albicans.

Green approach to synthesize novel thiosemicarbazide complexes, characterization; DNA-methyl green assay and in-silico studies against DNA polymerase

A thiosemicarbazide derivative as (E)-4-ethyl-1-(1-(naphthalen-1-yl) ethylidene) thiosemicarbazide (HAN) was synthesized then characterized to prepare [Co(HAN)Cl2·(H2O)2], [Ni(HAN)Cl2·(H2O)2]. H2O, and [Cd(HAN)Cl2] complexes. According to spectral and analytical data we could confirm the neutral bidentate mode of bonding via (C]S) and (C]N) groups to form 1:1 M ratio within the three complexes. The octahedral geometry was suggested for Co(II) and Ni(II) complexes according to electronic transitions assigned to 4T1g → 4T1g(P)(ʋ2) and 4T1g → 4A2g(F)(ʋ3) and 3A2g → 3T1g(P,υ3) and 3A2g → 3T1g(F,υ2), respectively. The values of nephelauetic ratio (β) in the ligand field parameters detect the ionic nature of new M-L bonds. The molecular ion peaks appeared in the mass spectra of two selected complexes confirming their molecular formulae. The conductometric study was performed for Cd(II) ion solution during variable additions of HAN to calculate association and formation constant of Cd(II)-HAN complex. DFT/B3LYP method was used to optimize the structures of the compounds and confirm the binding mode of the ligand. The distribution of C(5) = N(17) and C(13) = S(19) groups asserts their priority in coordination. Hirshfeld crystal properties were obtained via normalized contact distance (dnorm) and shape index in which the nitrogen atoms act as the best contact points in crystal packing. The biological screening was carried out against microbial strains as well as methyl green/DNA test. In vitro, the superiority of the ligand was clearly recorded in its biological effectiveness. In silico methods were implemented to confirm the activity of the ligand and to recognize the interaction features. The bioavailability, pharmacokinetics and drug-likeness were evaluated via Swiss-link. The data detect the ability of the ligand to penetrate barrier of brain (BBB) but not absorbed in gastrointestinal tract. Pharmit link and molecular docking were utilized to investigate the interaction of HAN with 1bna, 425d and 2k4l proteins. The best intercalation with protein pockets was observed with 2k4l protein, and searching the MolPort library detects a drug analog of MolPort-002-894-701. Finally, the results suggest the biological efficiency of the ligand, which may be asserted by specialists through intensive in-vivo studies.

Chemical and Pharmacological Prospection of the Ascidian Cystodytes dellechiajei.

Marine invertebrates are a traditional source of natural products with relevant biological properties. Tunicates are soft-bodied, solitary or colonial, sessile organisms that provide compounds unique in their structure and activity. The aim of this work was to investigate the chemical composition of the ascidian Cystodytes dellechiajei, selected on the basis of a positive result in biological screening for ligands of relevant receptors of the innate immune system, including TLR2, TLR4, dectin-1b, and TREM2. Bioassay-guided screening of this tunicate extract yielded two known pyridoacridine alkaloids, shermilamine B (1) and N-deacetylshermilamine B (2), and a family of methyl-branched cerebrosides (3). Compounds 2 and 3 showed selective binding to TREM2 in a dose-dependent manner. N-deacetylshermilamine B (2), together with its acetylated analogue, shermilamine B (1), was also strongly cytotoxic against multiple myeloma cell lines. TREM2 is involved in immunomodulatory processes and neurodegenerative diseases. N-deacetylshermilamine B (2) is the first example of a polycyclic alkaloid to show an affinity for this receptor.

High-Throughput Miniaturized Synthesis of PROTAC-Like Molecules.

The development of miniaturized high-throughput in situ screening platforms capable of handling the entire process of drug synthesis to final screening is essential for advancing drug discovery in the future. In this study, an approach based on combinatorial solid-phase synthesis, enabling the efficient synthesis of libraries of proteolysis targeting chimeras (PROTACs) in an array format is presented. This on-chip platform allows direct biological screening without the need for transfer steps. UV-induced release of target molecules into individual droplets facilitates further on-chip experimentation. Utilizing a mitogen-activated protein kinase kinases (MEK1/2) degrader as a template, a series of 132 novel PROTAC-like molecules is synthesized using solid-phase Ugi reaction. These compounds are further characterized using various methods, including matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) imaging, while consuming only a few milligrams of starting materials in total. Furthermore, the feasibility of culturing cancer cells on the modified spots and quantifying the effect of MEK suppression is demonstrated. The miniaturized synthesis platform lays a foundation for high-throughput in situ biological screening of potent PROTACs for potential anticancer activity and offers the potential for accelerating the drug discovery process by integrating miniaturized synthesis and biological steps on the same array.

Computational fragment-based drug design of potential Glo-I inhibitors

In this study, a fragment-based drug design approach, particularly de novo drug design, was implemented utilising three different crystal structures in order to discover new privileged scaffolds against glyoxalase-I enzyme as anticancer agents. The fragments were evoluted to indicate potential inhibitors with high receptor affinities. The resulting compounds were served as a benchmark for choosing similar compounds from the ASINEX® database by applying different computational ligand-based drug design techniques. Afterwards, the selection of potential hits was further aided by various structure-based approaches. Then, 14 compounds were purchased, and tested in vitro against Glo-I enzyme. Of the tested 14 hits, the biological screening results showed humble activities where the percentage of Glo-I inhibition ranged from 0–18.70 %. Compound 19 and compound 28, whose percentage of inhibitions are 18.70 and 15.80%, respectively, can be considered as hits that need further optimisation in order to be converted into lead-like compounds.

Integrating computational methods guided the discovery of phytochemicals as potential Pin1 inhibitors for cancer: pharmacophore modeling, molecular docking, MM-GBSA calculations and molecular dynamics studies.

Pin1 is a pivotal player in interactions with a diverse array of phosphorylated proteins closely linked to critical processes such as carcinogenesis and tumor suppression. Its axial role in cancer initiation and progression, coupled with its overexpression and activation in various cancers render it a potential candidate for the development of targeted therapeutics. While several known Pin1 inhibitors possess favorable enzymatic profiles, their cellular efficacy often falls short. Consequently, the pursuit of novel Pin1 inhibitors has gained considerable attention in the field of medicinal chemistry. In this study, we employed the Phase tool from Schrödinger to construct a structure-based pharmacophore model. Subsequently, 449,008 natural products (NPs) from the SN3 database underwent screening to identify compounds sharing pharmacophoric features with the native ligand. This resulted in 650 compounds, which then underwent molecular docking and binding free energy calculations. Among them, SN0021307, SN0449787 and SN0079231 showed better docking scores with values of -9.891, -7.579 and -7.097kcal/mol, respectively than the reference compound (-6.064kcal/mol). Also, SN0021307, SN0449787 and SN0079231 exhibited lower free binding energies (-57.12, -49.81 and -46.05kcal/mol, respectively) than the reference ligand (-37.75kcal/mol). Based on these studies, SN0021307, SN0449787, and SN0079231 showed better binding affinity that the reference compound. Further the validation of these findings, molecular dynamics simulations confirmed the stability of the ligand-receptor complex for 100ns with RMSD ranging from 0.6 to 1.8Å. Based on these promising results, these three phytochemicals emerge as promising lead compounds warranting comprehensive biological screening in future investigations. These compounds hold great potential for further exploration regarding their efficacy and safety as Pin1 inhibitors, which could usher in new avenues for combating cancer.

Bioactive monoterpenoids and acetophenones from the aerial parts of Eupatorium fortunei

Thirty-nine thymol and acetophenone derivatives, including eight pairs of enantiomers, were isolated from the aerial parts of Eupatorium fortunei. Their structures were assigned by detailed analyses of spectroscopic data and NMR calculations based on density functional theory, with 18 ones (1a/1b−14) being previously undescribed compounds. While the absolute configurations of 1a/1b, 2a/2b, 4, 6a/6b, 7, 11a/11b and 15a/15b−18a/18b were established by calculations of electronic circular dichroism data, that of 14 was determined by modified Mosher's method. Compounds 1a/1b and 2a/2b represent a previously unreported type of monoterpenoid dimers via an amide linkage, and compound 3 is a monoterpene-phenylpropanoid hybrid connected through an ester bond. Among the known molecules, the formerly mis-assigned structures of 15a/15b and 22 were revised, and pure natural enantiomers of 16a/16b−18a/18b were reported for the first time. Selective compounds showed antiradical and NO production inhibitory activities in the preliminary biological screening. Compound 31 was further demonstrated to alleviate oxidative stress by activating Nrf2 signaling pathway.

Discovery, Identification, and Mode of Action of Phenolics from Marine-Derived Fungus Aspergillus ustus as Antibacterial Wilt Agents.

The bacterial wilt caused by Ralstonia solanacearum seriously affects crop yield and safety and is difficult to control. Biological activity-guided screening led to the isolation of 11 phenolic compounds including three undescribed compounds (carnemycin H-I and stromemycin B) from the secondary metabolites of a marine-derived Aspergillus ustus. One new compound is an unusual phenolic dimer. Their structures were elucidated by comprehensive spectroscopic data and J-based configurational analysis. The antibacterial activities of the isolated compounds against R. solanacearum were evaluated. Compound 3 exhibited excellent inhibitory activity with an MIC value of 3 μg/mL, which was comparable to that of streptomycin sulfate. Additionally, 3 significantly changed the morphology and inhibited the activity of succinate dehydrogenase (SDH) to interfere with the growth of R. solanacearum. Molecular docking was conducted to clarify the potential mechanisms of compound 3 with SDH. Further in vivo experiments demonstrated that 3 could remarkably inhibit the occurrence of bacterial wilt on tomatoes.

Trifurcated Splitting of Water Droplets on Engineered Lithium Niobate Surfaces.

Controlled splitting of liquid droplets is a key function in many microfluidic applications. In recent years, various methodologies have been used to accomplish this task. Here, we present an optofluidic technique based on an engineered surface formed by coating a z-cut iron-doped lithium niobate crystal with a lubricant-infused layer, which provides a very slippery surface. Illuminating the crystal with a light spot induces surface charges of opposite signs on the two crystal faces because of the photovoltaic effect. If the light spot is sufficiently intense, millimetric water droplets placed near the illuminated spot split into two charged fragments, one fragment being trapped by the bright spot and the other moving away from it. The latter fragment does not move randomly but rather follows one of three well-defined trajectories separated by 120°, which reflect the anisotropic crystalline structure of Fe:LiNbO3. Numerical simulations explain the behavior of water droplets in the framework of the forces induced by the interplay of pyroelectric, piezoelectric, and photovoltaic effects, which originate simultaneously inside the illuminated crystal. Such a synergetic effect can provide a valuable feature in applications that require splitting and coalescence of droplets, such as chemical microreactors and biological encapsulation and screening.

Biological screening for congenital toxoplasmosis in newborns from Jataí, Goiás, Brazil: a cross-sectional study.

To conduct a serological screening for toxoplasmosis in the heel prick test and to evaluate its epidemiological aspects in newborns and postpartum women in Jataí, Goiás. Cross-sectional epidemiological study for the biological screening of newborns in Jataí, Goiás. The study participants amounted to 228 newborns, whose samples were collected between the third and seventh day of life. IgG antibodies against Toxoplasma gondii were detected in 40.79% (93/228) of the samples; out of these, 23.6% (22/93) had high IgG antibody titers, leading to the collection of two other peripheral blood samples and the detection of a decrease in these titers. The findings show the importance of strengthening actions in primary health care to prevent infection and training health professionals in this area to equip them with information regarding cases of reinfection and reactivation of infection in pregnant women, minimizing risks for babies.

Highly efficient, catalyst-free, one-pot sequential four-component synthesis of novel spiroindolinone-pyrazole scaffolds as anti-Alzheimer agents: in silico study and biological screening.

Alzheimer's disease is a neurodegenerative disorder that impacts memory, thinking, and behavior, and currently, there is no effective cure available for its treatment. This study explored a one-pot strategy for synthesizing spiroindolinone-pyrazole derivatives through a sequential four-component condensation reaction. These derivatives were further investigated for their potential as anti-Alzheimer's disease agents. The developed synthetic procedure provides remarkable advantages, including a clean reaction profile, abundant starting materials, operational simplicity, and easy purification without traditional methods with good to excellent yields (84-96%). Next, the biological potencies of the newly synthesized spiroindolinone-pyrazole derivatives against AChE and BChE as Alzheimer's disease-related targets were determined. Also, the kinetic study and cytotoxicity of the most potent derivative were investigated. Furthermore, molecular docking and molecular dynamics evaluations were performed employing in silico tools to investigate the interaction, orientation, and conformation of the potent analog over the active site of the enzyme.

Mixed Ligand Complexes: Room Temperature Synthesis, Structure Determination and Biological Utility

The synthesis of mixed ligand complexes at room temperature involving a Schiff base derived from a heterocyclic compound (S5) and 5-chloro-8-hydroxyquinoline ligands, utilizing transition metals such as Fe(III), Co(II), Ni(II), and Cu(II), has been documented. The formation of the Schiff base (S5) is achieved by reacting 2-hydroxy-3-methoxy-5-nitrobenzaldehyde with 2-aminobenzothiazole in ethanol, with glacial acetic acid serving as a catalyst. Various analysis techniques utilized to analyze the obtained compounds for their structure determination. Biological screening results revealed the obtained metal-ligand combinations are possible candidates for use as antibacterial, antimalarial and antioxidant agents owing to their potential activities.

Practical synthesis of isoindolines yields potent colistin potentiators for multidrug-resistant Acinetobacter baumannii.

An efficient and practical one-pot synthesis of isoindolines from readily available starting materials was achieved under mild conditions by implementing an isoindole umpolung strategy. A variety of isoindolines were prepared with good to excellent yields. Biological screens of these identified compounds demonstrated that they are potent potentiators of colistin for multi-drug resistant Acinetobacter baumannii.

Novel pyrimidine linked acyl thiourea derivatives as potent α-amylase and proteinase K inhibitors: design, synthesis, molecular docking and ADME studies.

To discover promising therapeutic agents, novel diaryl pyrimidine linked acyl thiourea derivatives (6a-j) were designed and synthesized via straightforward and multistep synthesis. The structure of these derivatives (6a-j) was confirmed by FTIR, 1H, and 13C NMR spectroscopic techniques. The in vitro biological screening of these compounds was carried out to assess their bacterial, α-amylase, and proteinase K inhibition potential. The results manifested that the developed molecules (6a-j) possessed a remarkable inhibition potential against targeted α-amylase and proteinase K enzymes. The compounds 6j and 6g were found to be the most potent α-amylase inhibitors with IC50 values of 1.478 ± 0.051 and 1.509 ± 0.039 μM, respectively. Meanwhile, the compounds 6a, 6f, and 6e having IC50 values of 1.790 ± 0.079, 1.794 ± 0.080, and 1.795 ± 0.080 μM, respectively, showed high proteinase K inhibitory activity. A moderate antibacterial activity is also displayed by these compounds (6a-j). The different substitution on the framework of pyrimidine linked acyl thiourea pharmacophore, provided the valuable basis for structure-activity relationship studies. Additionally, to identify the binding affinities of our desired compounds, molecular docking study was used. ADME analysis was also conducted to explore the physicochemical properties. Hence, these studies shed light on the significance of pyrimidine-based acyl thiourea to attain potent efficacy in drug discovery.

N-Substituted dihydropyridines as promising EGFR tyrosine kinase inhibitors against breast cancer: Design, synthesis, biological evaluation, docking, and molecular dynamics simulations

Breast cancer is a major health concern, chemotherapy and radiotherapy face ongoing challenges due to drug resistance and toxicity. EGFR is a key driver of tumorigenesis, and its amplification or mutation is common in breast cancer. Tyrosine kinase inhibitors (TKIs), such as erlotinib and lapatinib, inhibit the EGFR pathway by competitively binding to the adenosine triphosphate pocket of EGFR. The Hantzsch reaction is a versatile method for synthesizing dihydropyridine derivatives that exhibit a range of pharmacological effects. This study aims for the molecular design, synthesis, analysis, and biological screening of N-Substituted Hantzsch dihydropyridines for their potential as EGFR inhibitors. The results showed that synthesized compounds inhibited the proliferation of MCF-7 breast cancer cells and modulated the expression of genes involved in the PI3K/Akt signaling pathway, which is downstream events of EGFR. Among all the synthesized compounds, NHD-3, NHD-4, and NHD-5 displayed the most potent inhibitory activity against MCF-7 cells with IC50 value of 17.24 ± 1.23, 10.92 ± 1.03, and 07.34 ± 0.86 µM, respectively. The potent dihydropyridines derivatives were also screened for EGFRwt inhibition assay and among the three compounds tested, NHD-5 showed reasonably good inhibitory activity with an IC50 value of 20.10 ± 1.30 nM compared to a Lapatinib with IC50 value of 10.28 ± 1.01 nM. Molecular docking and molecular dynamics simulations were performed to provide insight into the binding modes, interactions, and stability of these compounds with EGFR.