Human Estrogen Receptor Alpha Research Articles

Activities of Chalcone Derivatives from Boesenbergia rotunda Against Human Estrogen Receptor Alpha of Breast Cancer by In Silico

The high prevalence of cancer must be overcome with prompt and appropriate prevention and treatment. New drug design is an effort to develop existing drugs, and their molecular structure and biological activity have been known through structural modification. It encourages researchers to explore Indonesia's natural resources, especially plants with anticancer activity, namely by synthesizing chalcone-derived compounds derived from the isolation of Fingerroot rhizomes (Boesenbergia rotunda). The most common flavonoid compound found in rhizomes fingerroot plants is pinostrobin. Pinostrobin compounds and their derivatives are synthesized, resulting in chalcone compounds and their derivative modifications. The author conducted an in-silico test on pinostrobin compounds and 19 of their derivatives, chalcone compounds, and 18 derivatives using estrogenic-a receptors with PDB codes 3ERD and 1G50. The author hoped that from this silico test, compounds with more potential as anticancer for breast cancer would be obtained based on the results of docking with 3ERD and 1G50 receptors and can then be synthesized. In the results of this study, the compounds Bis-4-bromobenzyoxychalcone and Bis-4-chlorobenzyloxychalcone are the most appropriate compounds to be synthesized. It is hoped that in the future, they can be continued with activity tests of these compounds, both in vitro and in vivo, because these compounds are predicted to have the best activity and do not have hepatotoxic or other toxicity effects

Exploring compound suitability and employing DFT calculations, molecular docking, and dynamics simulation to investigate potent compounds from podophyllum medicinal plants for breast cancer therapy

BackgroundBreast cancer, one of the most often diagnosed malignancies worldwide, continues to take countless women's lives. Its treatment usually involves targeting the human estrogen receptor alpha (ERα). Current research explores the potential of natural compounds to regulate ERα activity, providing a hopeful direction for breast cancer therapy. Our study utilized a comprehensive approach to identify promising natural compounds for breast cancer treatment, including quantum descriptors, molecular docking, molecular dynamics simulations, and ADMET/pharmacokinetics analysis. ResultsSix natural compounds derived from podophyllum medicinal plants, namely 4-demethylpodophyllotoxin (NP1), α-peltatin (NP2), podophyllotoxin (NP3), deoxypodophyllotoxin (NP4), podophyllotoxone (NP5), and β-peltatin (NP6), were investigated as potential selective estrogen receptor α (ERα) inhibiting agents for breast cancer. These compounds demonstrated the strongest binding affinity to the target enzyme, with binding energies of − 8.9 and − 8.1 kcal/mol, respectively. Further assessments of drug-likeness and ADME properties were conducted for these compounds, along with quantum calculations (HOMO–LUMO) to evaluate their reactivity. Additionally, molecular dynamics studies were performed to assess the stability of the NP1 and NP2 protein–ligand complexes.ConclusionsWe analyzed six natural compounds comprehensively, evaluating their ADME properties, molecular docking interactions, quantum descriptors, and dynamic simulations. Our findings demonstrate that these natural compounds are promising possibilities for treating breast cancer. Additionally, they may provide a basis for developing future compounds targeting estrogen receptor α (ERα) activity.Graphical abstract

Human estrogen receptor alpha (ERα) targeted cyclic peptides inhibit cell growth and induce apoptosis in MCF-7 cells

Abstract Objectives Human estrogen receptor alpha (ERα) is considered an important target, especially in the treatment of breast cancer, as it has a vital role in cancer development. ERα-targeted therapies generally target the ligand binding domain (LBD) of ERα. However, over time, cells develop resistance to this mechanism alternative approaches to inhibit ERα activity target ERα–DNA or ERα–cofactor interactions. Inhibitors of ERα–cofactor interactions are designed by targeting the hydrophobic hollow region of the receptor box LXXLL motif. Methods In this context, helix-stabilized cyclic peptides (SPs) designed with in silico approaches were obtained by solid phase peptide synthesis. The effects of SPs on MCF-7 cells were examined with MTT and ATP, and qPCR and flow cytometry were used for further analysis. Results Our results demonstrated that the SPs were effective only in MCF-7 cells expressing ERα. In addition, cyclic peptide combinations (SPCs) showed anti-proliferative and toxic effects on MCF-7 cells. The impact of SPCs with the highest inhibitory effect in MCF-7 cells on ERα-related genes and markers of apoptosis was revealed. Moreover, the flow cytometry analysis result used to examine apoptotic cells proved the apoptosis of SPCs in MCF-7 cells. Conclusions These findings suggest that our novel SPs, which inhibit coactivator interactions of ERα, induce apoptosis of MCF-7 cells. Thus, considering this strong effect of SPs in the inhibition of receptors, it is pointed out that they can be further developed as an alternative to current clinical treatments or as an auxiliary approach in the generating of new targeted peptide-based therapies.

Methylimidazolium ionic liquids - A new class of forever chemicals with endocrine disrupting potential

A class of chemical with a potentially important perceived future contribution to the net zero carbon goal (as "green" solvents) is the methylimidazolium ionic liquids (MILs). These solvents are used in industrial processes such as biofuel production yet little is known about their environmental stability or toxicity in man although one MIL - 1-octyl-3-methylimidazolium (M8OI) - has been shown to activate the human estrogen receptor alpha (ERα). The stabilities of the chloride unsubstituted methylimidazolium (MI) and MILs possessing increasing alkyl chain lengths (2C, 1-ethyl-3-methylimidazolium (EMI); 4C, 1-butyl-3-methylimidazolium (BMI); 6C; 1-hexyl-3-methylimidazolium (HMI), 8C, M8OI; 10C, 1-decyl-3-methylimidazolium (DMI)) were examined in river water and a human liver model system. The MILs were also screened for their abilities to activate the human ERα in vitro and induce uterine growth in pre-pubertal rats in vivo. Short chain MILs (EMI, BMI and HMI) underwent negligible metabolism and mineralisation in river water; were not metabolised in a model of human liver metabolism; activated the human ERα in vitro and were estrogenic in vivo in rats. A structure-based computational approach predicted short chain MIL binding to both the estrogen binding site and an additional site on the human estrogen receptor alpha. Longer chain MILs (M8OI and DMI) were metabolised in river water and partially mineralised. Based on structure-activity considerations, some of these environmentally-derived metabolites may however, remain a hazard to the population. MILs therefore have the potential to become forever chemicals with adverse effects to both man, other animals and the environment in general.

Enhanced eMAGE applied to identify genetic factors of nuclear hormone receptor dysfunction via combinatorial gene editing

Technologies that generate precise combinatorial genome modifications are well suited to dissect the polygenic basis of complex phenotypes and engineer synthetic genomes. Genome modifications with engineered nucleases can lead to undesirable repair outcomes through imprecise homology-directed repair, requiring non-cleavable gene editing strategies. Eukaryotic multiplex genome engineering (eMAGE) generates precise combinatorial genome modifications in Saccharomyces cerevisiae without generating DNA breaks or using engineered nucleases. Here, we systematically optimize eMAGE to achieve 90% editing frequency, reduce workflow time, and extend editing distance to 20 kb. We further engineer an inducible dominant negative mismatch repair system, allowing for high-efficiency editing via eMAGE while suppressing the elevated background mutation rate 17-fold resulting from mismatch repair inactivation. We apply these advances to construct a library of cancer-associated mutations in the ligand-binding domains of human estrogen receptor alpha and progesterone receptor to understand their impact on ligand-independent autoactivation. We validate that this yeast model captures autoactivation mutations characterized in human breast cancer models and further leads to the discovery of several previously uncharacterized autoactivating mutations. This work demonstrates the development and optimization of a cleavage-free method of genome editing well suited for applications requiring efficient multiplex editing with minimal background mutations.

The physiological and biochemical basis of potency thresholds modeled using human estrogen receptor alpha: implications for identifying endocrine disruptors

The endocrine system functions by interactions between ligands and receptors. Ligands exhibit potency for binding to and interacting with receptors. Potency is the product of affinity and efficacy. Potency and physiological concentration determine the ability of a ligand to produce physiological effects. The kinetic behavior of ligand-receptor interactions conforms to the laws of mass action. The laws of mass action define the relationship between the affinity of a ligand and the fraction of cognate receptors that it occupies at any physiological concentration. We previously identified the minimum ligand potency required to produce clinically observable estrogenic agonist effects via the human estrogen receptor-alpha (ERα). By examining data on botanical estrogens and dietary supplements, we demonstrated that ERα ligands with potency lower than one one-thousandth that of the primary endogenous hormone 17β-estradiol (E2) do not produce clinically observable estrogenic effects. This allowed us to propose a Human-Relevant Potency Threshold (HRPT) for ERα ligands of 1 × 10–4 relative to E2. Here, we test the hypothesis that the HRPT for ERα arises from the receptor occupancy by the normal metabolic milieu of endogenous ERα ligands. The metabolic milieu comprises precursors to hormones, metabolites of hormones, and other normal products of metabolism. We have calculated fractional receptor occupancies for ERα ligands with potencies below and above the previously established HRPT when normal circulating levels of some endogenous ERα ligands and E2 were also present. Fractional receptor occupancy calculations showed that individual ERα ligands with potencies more than tenfold higher than the HRPT can compete for occupancy at ERα against individual components of the endogenous metabolic milieu and against mixtures of those components at concentrations found naturally in human blood. Ligands with potencies less than tenfold higher than the HRPT were unable to compete successfully for ERα. These results show that the HRPT for ERα agonism (10–4 relative to E2) proposed previously is quite conservative and should be considered strong evidence against the potential for disruption of the estrogenic pathway. For chemicals with potency 10–3 of E2, the potential for estrogenic endocrine disruption must be considered equivocal and subject to the presence of corroborative evidence. Most importantly, this work demonstrates that the endogenous metabolic milieu is responsible for the observed ERα agonist HRPT, that this HRPT applies also to ERα antagonists, and it provides a compelling mechanistic explanation for the HRPT that is grounded in basic principles of molecular kinetics using well characterized properties and concentrations of endogenous components of normal metabolism.

Pinostrobin and its derivatives as novel anti-breast cancer agents against human oestrogen receptor alpha: In silico studies of ADMET, docking, and molecular dynamics

Background: Breast cancer is the main cause of cancer death among women. The issue is more complex due to the side effects and resistance of the currently used breast cancer drugs. Pinostrobin, a compound found in Boesenbergia pandurata, has anticancer activity. Modifying the structure of pinostrobin can improve drug bioavailability, reduce toxicity, and work more selectively. Objective: This study aimed to predict the potentials of pinostrobin and its derivatives as anti-breast cancer agents against human oestrogen receptors by an in silico approach. Methods: The pkCSM was utilised to predict the ADMET characteristics. The interaction of ligands with the binding site of the human oestrogen receptor alpha with PDB ID 3ERT was used for molecular docking using MOE, and AMBER was then used for molecular dynamic simulation. Results: Pinostrobin pentanoate had good pharmacokinetic properties and was neither mutagenic nor hepatotoxic. Based on molecular docking, it was more potent compared to pinostrobin, with binding free energy values of -7.849 kcal/mol. Furthermore, in the interaction stability evaluation using 100 ns molecular dynamic simulation by the MM-GBSA calculation method, pinostrobin pentanoate had a stable interaction with a total bond energy value of -9.943 kcal/mol. Conclusion: Pinostrobin pentanoate has the potential as an anti-breast cancer alternative through the human oestrogen receptor alpha.

Molecular Dynamic Simulation and 3d-pharmacophore Modeling of Alpha Mangostin and Its Derivatives against Estrogen Alpha Receptor

Background: Human estrogen receptor alpha (ERα), which is known to play a role in mediating cell proliferation, metastasis, and resistance to apoptosis, is one of the targets of breast cancer therapies. Alpha mangostin (AM) is an active xanthone compound from Garcinia mangostana L. which has activity as an ERα inhibitor. Objectives: This research aims to predict the pharmacokinetic and toxicity, and to study the molecular interactions of AM derivatives with the ERα using computer-aided simulation approaches through molecular docking, molecular dynamic, and pharmacophore screening to develop novel anti-breast cancer agents. Methods: Marvinsketch and Chimera programs were used to design and optimize the structure of AM and its derivatives. For screening the pharmacokinetic and toxicity profiles, the PreADMET web was used. The AutoDockTools 1.5.6 and LigandScout 4.4.3 Advanced software were used to conduct the molecular docking simulation and pharmacophore screening, respectively, while the molecular dynamic simulation was performed using AMBER 16. The results were visualized by Biovia Discovery Studio. Results: Molecular docking using Autodock showed that FAT10 derivate has lower binding free energy (ΔG) (-12.04 kcal/mol) than AM (-8.45 kcal/mol) when docking to ERα and both performed the same hydrogen bond with Thr347. These support the results of the MMPBSA calculation on dynamic simulation which shows FAT10 (-58.4767 kcal/mol) has lower ΔG than AM (-42.7041 kcal/mol) and 4-OHT (- 49.0821 kcal/mol). The pharmacophore screening results also showed that FAT10 fitted the pharmacophore with a fit score of 47.08. Conclusion: From the results, it can be suggested that FAT10 has promising activity as ERα antagonist. Further in vitro and in vivo experiments should be carried out to support these in silico studies.

Phytochemical Characterization and Anticancer Screening of Cladophora goensis and Cladophora glomerata Extracts

Background Cladophora goensis and Cladophora glomerata has been widely identified in the Indian Oceans. Marine algae have been identified as potential pharmacological agents useful in the treatment of diabetes, hypertension, infections, and cancers. Objectives The present study aimed to evaluate the phytochemicals in C. goensis and C. glomerata using gas chromatography-mass spectroscopy (GC-MS), and to study the molecular interactions with cancer-related proteins using molecular docking techniques. Materials and methods GC-MS analysis was done using electron impact ionization at 70eV and the data was evaluated using total ion count for compound identification and quantification. AutoDock 4.0 version was used for the molecular docking analysis. Results The methanolic extracts of algae when subjected to GC-MS analysis, 19 compounds from C. goensis and 11 phytocompounds from C. glomerata were identified. The significant molecular interactions of phytochemicals of C. goensis (6-nitro-3 H-quinazolin-4-one, Isoquinoline, 1,2,3,4-tetrahydro-7-methoxy-2-methyl-8-(phenyl methoxy) and 9-Decen-1-ol, pentafluropropionate) and C. glomerata (phytol, palmitic acid, and octadec-9-enoic acid) against human epidermal growth factor receptor (4WRG), poly (ADP-ribose) polymerase-1 (4UND), human estrogen receptor alpha ligand-binding domain (3ERT), human peroxisome proliferator-activated receptor alpha ligand-binding domain (3VI8), and human topoisomerase (1EJ9) have been demonstrated. Conclusion The phytochemicals of methanolic extracts of C. goensis and C. glomerata showed potential interactions with cancer-related proteins.

Synthesis, Structure, and In Vitro Pharmacological Evaluation of some New Pyrimidine-2-Sulfonamide Derivatives and Their Molecular Docking Studies on Human Estrogen Receptor Alpha and CDK2/Cyclin Proteins

Synthesis, Structure, and In Vitro Pharmacological Evaluation of some New Pyrimidine-2-Sulfonamide Derivatives and Their Molecular Docking Studies on Human Estrogen Receptor Alpha and CDK2/Cyclin Proteins

Molecular Docking Studies and In-silico ADMET Profile Analysis of Triphala Plant constituents Morin and 9, 10-anthraquinone as Potential Inhibitors of human Estrogen Receptor Alpha

The expanding global burden of cancer demands novel treatment options. Herbal medicine offers a viable alternative to conventional cancer treatment. Breast cancer is frequently diagnosed cancer among women all over the world. Normal breast cells and specific breast cancer cells containestrogen and progesterone receptors. Estrogen and progesterone bind to receptors and stimulate cancer cell proliferation and expansion with growth factors (e.g., oncogenes and mutated gene suppressors). Tamoxifen, raloxifene, andtoremifene are among the most frequently used drugs for breast cancer, whereas estrogen continues to be produced in breast cancer cells. These medications primarily function against theestrogen binding to theestrogenreceptors on these cells. Protein-ligand interaction plays a major role in structural drug design. In this study, the molecular interactions between theHuman estrogen receptor alpha (PDB ID: 2IOK), and the two ligands isolated from Triphala Ayurvedic formulation morin, and 9,10-anthraquinone, an endogenous estrogen receptor ligand, estradiol, and the three economically available breast cancer drug, tamoxifen, raloxifene, and toremifene using the AutodockVina software tool. The ADMET properties of these substances were determinedusing popular web-based software tools that include preADMET, admetSAR, Molinspiration, and SwissADME.The present available anti-cancer drugs have so many side effects. The present work is aimed at determining the efficacy ofestrogen receptor inhibitors and the safety profile of morin, anthraquinone molecules isolated from Triphalapolyherbal formulation by in-silico methods. The binding energies (Kcal/mol) of ligands with human estrogen receptor were calculated as follows: morin (-9.0), 9,10-anthraquinone (-8.7), estradiol (-10.1), tamoxifen (-9.6), raloxifene (-9.8) and toremifene (-8.9). The resultssupported the drug-like properties of the molecules tested and they are likely to have a therapeutic effect.Further,in vivo and pre-clinical trials of the most active compound are also worthwhile for producing effective inhibitors.

Novel hybrid motifs of 4-nitroimidazole-piperazinyl tagged 1,2,3-triazoles: Synthesis, crystal structure, anticancer evaluations, and molecular docking study

4-((4-(1-benzyl-2-methyl-4-nitro-1H-imidazole-5-yl)piperazine-1-yl)methyl)-1-substituted-1H-1,2,3-triazole motifs are designed and synthesized via click chemistry. The reaction of 1-(N1-benzyl- 2-methyl-4-nitro-1H-imidazole- 5-yl)-4-(prop-2-yn-1-yl) piperazine 5 as new scaffold with diverse primary azides to selectively produce 1,4-disubstituted-1,2,3-triazoles 9a-k, 10a-c and 11a-q. Physicochemical methods: when 1H NMR, 13C NMR, and HRMS are utilized to fully characterize all synthesized compounds. X-ray structural determination and analysis for compound 9a is also performed. The newly designed chromophores are assessed for their anti-proliferative potency against three selected human cancer cell lines (MCF-7, HepG2, and PC3), and one normal cell line (Dermal/Fibroblast). Compounds 9g and 9k have shown potent activities against the MCF-7cell line with IC50 values of (2.00±0.03μM) and (5.00±0.01μM) respectively. ADMET studies and Molecular docking investigations are performed on the most active hybrid nitroimidazole derivatives 9g and 9k with 4-hydroxytamoxifen (4-OHT) at the human estrogen receptor alpha (hER) during binding active sites to study the ligand-protein interactions and free binding energies at atomic levels. The triazole ring in the 9g derivative forms a hydrogen bond with Asp58 with distance 3.2Å. And it is found that polar contact with His231 amino acid residue. In silico assessment of the compounds showed very good pharmacokinetic properties based on their physicochemical values, also the ADMET criteria of the most active hybrid systems are within the acceptable range.

Investigating Endocrine Disrupting Impacts of Nine Disinfection Byproducts on Human and Zebrafish Estrogen Receptor Alpha.

Disinfection byproducts (DBPs) cause endocrine disruption via estrogenic or anti-estrogenic effects on estrogen receptors. However, most studies have focused on human systems, with little experimental data being presented on aquatic biota. This study aimed to compare the effects of nine DBPs on zebrafish and human estrogen receptor alpha (zERα and hERα). In vitro enzyme response-based tests, including cytotoxicity and reporter gene assays, were performed. Additionally, statistical analysis and molecular docking studies were employed to compare ERα responses. Iodoacetic acid (IAA), chloroacetonitrile (CAN), and bromoacetonitrile (BAN) showed robust estrogenic activity on hERα(maximal induction ratios of 108.7%, 50.3%, and 54.7%, respectively), while IAA strongly inhibited the estrogenic activity induced by 17β-estradiol (E2) in zERα (59.8% induction at the maximum concentration). Chloroacetamide (CAM) and bromoacetamide (BAM) also showed robust anti-estrogen effects in zERα (48.1% and 50.8% induction at the maximum concentration, respectively). These dissimilar endocrine disruption patterns were thoroughly assessed using Pearson correlation and distance-based analyses. Clear differences between the estrogenic responses of the two ERαs were observed, whereas no pattern of anti-estrogenic activities could be established. Some DBPs strongly induced estrogenic endocrine disruption as agonists of hERα, while others inhibited estrogenic activity as antagonists of zERα. Principal coordinate analysis (PCoA) showed similar correlation coefficients for estrogenic and anti-estrogenic responses. Reproducible results were obtained from computational analysis and the reporter gene assay. Overall, the effects of DBPs on both human and zebrafish highlight the importance of controlling their differences in responsiveness for estrogenic activities including the water quality monitoring and endocrine disruption, as DBPs have species-specific ligand-receptor interactions.

Synthesis, Cytotoxicity, Molecular Docking and ADME Assay of Novel Morpholine appended 1,2,3‐Triazole Analogues

AbstractSynthesis of novel 4‐(1‐(2‐cyclopropylphenyl)‐2‐(4‐substituted‐1H‐1,2,3‐triazol‐1‐yl)ethyl)morpholine analogues were demonstrated by conventional synthetic procedures. The structure of all the newly synthesized compounds were determined by 1H‐NMR, 13C‐NMR, HRMS and CHN analysis. Cytotoxicity of all synthesized compounds were tested against MCF‐7 Cell lines in different concentrations. The IC50 value of compounds was calculated using graph Pad Prism Version5.1. 4‐chloro substituted analogue exhibited superior result than the standard reference Doxorubicin drug. Compounds which are containing 3‐methoxy, 2‐methoxy and 4‐methoxy substituents showed a moderate effect, and 2‐chloro, 3‐triflouromethyl and 2‐methyl substituted analogues showed lower results. Molecular docking studies performed on the crystal structure of human estrogen receptor alpha ligand‐binding domain with all molecules and are well in agreement with cell line studies. The predicted pharmacokinetics support that these compounds have more drug‐likeness properties.

Azole pesticide products and their hepatic metabolites cause endocrine disrupting potential by suppressing the homo-dimerization of human estrogen receptor alpha

We selected azole pesticides products that are managed by setting maximum residue limits (MRLs) in the Republic of Korea and describe the estrogen receptor (ER) α-related negative effect to endocrine system using in vitro Organization for Economic Cooperation and Development performance-based test guideline. No azoles were found to be an ERα agonist. Conversely, three azoles (bitertanol, cafenstrole, and tebufenpyrad) were determined to be ERα antagonists. In addition, the ERα antagonistic activities of bitertanol, cafenstrole, and tebufenpyrad were not significantly perturbed in the existence of phase I (hydroxylation, dealkylation, oxidation or reduction) and phase II (conjugation). Regarding the mechanism underlying their ERα-mediated endocrine disrupting potentials, ERα proteins cannot be translocated to the nucleus by suppressing the dimerization of ERα in the cytoplasm by bitertanol, cafenstrole, and tebufenpyrad. These data indicated that azole pesticide products show the capability to interfere the ERα-related human endocrine system. Furthermore, we identified the mechanism of ERα-mediated endocrine disrupting by azole insecticide products through this study.

Pharmacophore Modeling and Molecular Docking of Flavonoid Derivatives in Abelmoschus manihot Against Human Estrogen Receptor Alpha of Breast Cancer

Tamoxifen is the most commonly used anti-estrogen adjuvant therapy for estrogen receptor-positive breast cancer. However, it is associated with an increased risk of some serious side effects, such as uterine cancer, stroke, and pulmonary embolism. The flavonoid compounds in the leaves of A. manihot inhibited the growth of 4T1 breast cancer cells at a CTC50 concentration of 185.06 μg/ml. Therefore, this study aims to examine the molecular interactions and pharmacophore modeling based on the interaction of 4-OHT with human ER, followed by the molecular docking of the flavonoid derivatives with human ERα. The molecular docking simulations and 3D structure-based pharmacophore models were used to identify the molecular interactions of flavonoid derivatives in A. manihot on estrogen receptors (ERα) (PDB ID: 3ERT). The results showed that the binding energies of the flavonoid derivatives in isorhamnetin and isoquercitrin were -8.68 kcal/mol and -8.75 kcal/mol, respectively. This compound also interacted with Arg394 and Glu353 important amino acid residues in the ERα-binding pocket. Meanwhile, the pharmacophore fit scores of isorhamnetin and isoquercitrin were 82.36% and 84.91%, respectively. The flavonoid derivatives in A. manihot had pharmacophore fit resulting from the 4-OHT complex with ER, and therefore they had potential as ERα antagonists. Out of the 10 flavonoid derivatives, isorhamnetin and isoquercitrin showed the best docking scores and could be used as candidates for new anti-breast cancer drugs with antagonistic activity against ERα.

Generation and Characterization of Novel Diagnostic Mouse Monoclonal Antibodies Against Human Estrogen Receptor Alpha and Progesterone Receptor.

Estrogen and progesterone regulate the growth and development of several human cells and tissues. Their corresponding receptors (ER and PR) are important diagnostic and prognostic indicators for cancers of the breast and reproductive organs. Immunohistochemical analysis of ER and PR is the current standard method for evaluating the expression of these receptors in different cancers. Nonetheless, there is a significant lack of reproducibility of IHC results in laboratories worldwide, necessitating to develop more sensitive and specific antibodies for ER and PR IHC staining. ER and PR-specific monoclonal antibodies (MAbs) were generated by immunizing mice with synthetic peptides from ERα and PR. The isotypes and affinity constants of the selected MAbs were determined, and their specificities were assessed by peptide-specific ELISA, IHC, Western-blot analysis, and flow cytometry. In addition, the reactivity of generated MAbs was compared with that of the commercially-available anti-ER and anti-PR antibodies in IHC using normal and cancerous tissue sections. Moreover, 200 breast cancer tissue samples were stained using the newly generated MAbs along with commercial antibodies by IHC, and the sensitivity, specificity and accuracy of our MAbs were evaluated. Among different MAbs generated in this study, two anti-ER and one anti-PR MAbs specifically detected the target antigens in normal and cancerous tissues in IHC. Further analyses confirmed the specificity of the MAbs in Western blotting and flow cytometry using a panel of ER and PR positive cell lines. The sensitivity, specificity and accuracy calculated for clone 1B9 (anti-ER) were 92.3%, 94.8% and 93%, and for clone 3D6 (anti-PR) were 93.0%, 94.3% and 93.5%, respectively. Our novel anti-ER and PR MAbs could be considered as suitable tools for diagnostic and research purposes.

Design, synthesis, bioevaluation, DFT, docking, and molecular dynamic simulation for selected novel 1,3,4-Oxadiazole - indole derivatives hybrid against estrogen receptor alpha

• The novel method was developed to synthesize newly designed 1,3,4-Oxadiazole incorporated N1-Indole derivatives. • The screening results of the synthesized compounds against cancer cell lines have shown antitumor properties; among these compounds, the compound 3d has shown promising antitumor properties against Triple Negative Breast cancer cell lines, a potent molecule which displayed IC50 values of 10.56, and 22.61µM against MDA MB 468, and MDA MB 231 cell lines respectively. • In vitro antibody specific for Human Estrogen Receptor alpha assay showed an IC50 value of 5.27 µM for 3d compound. • The computation biology studies for synthesized compounds have shown that, the compound 3d is the potential candidate against estrogen receptor alpha. Breast tumors overexpress ER alpha and it initiates, as well as the progression of breast cancer. Studies have proven that anti-estrogen therapy (tamoxifen) is used to treat breast cancer. For the first time, in vitro bio-evaluation, molecular docking, and DFT simulations were used to investigate the essential interactions of the estrogen receptor (ER) alpha with selected novel 1,3,4-Oxadiazole - indole derivatives hybrid against Triple Negative Breast Cancer (TNBC) cell lines. The key step of this synthesis is the oxidative cyclization of N-acylhydrazones, 2(a-j), in 20 min using reagent BTI [bis(trilfuoroacetoxy)iodobenzene]. Synthesized compounds, 3(a-j), were screened against a series of cancer cell lines, and among these compounds, the compound, 3d((5’-Bromo-1H-indol-1-yl) methyl)-5’’-(3-bromo-4-methylphenyl)-1,3,4-Oxadiazole, as a potent molecule which displayed IC 50 values of 10.56 and 22.61µM against MDA MB 468, and MDA MB 231 cell lines respectively. In vitro antibody specific for Human Estrogen Receptor alpha assay showed an IC 50 value of 5.27 µM for compound 3d. The structures were optimized to their molecular geometry (3d) by the DFT method with B3LYP hybrid functional and 6-311++g(d,p) basis set. The electrostatic potential map was plotted to understand the energy distribution and chemical reactive regions of the molecule. The optimized structures were used to perform in silico studies. Molecular docking followed by molecular dynamics trajectories is essential to investigate the stable binding of 3d to estrogen receptor alpha. Overall, results show that 3d has the potential to be further explored for the treatment of metastatic breast cancer.

Experimental, DFT study, and in silico molecular docking investigations of dichlorodiphenyltrichloroethane against human estrogen receptor alpha

Advanced computational tools allowed to study a pure commercial sample of dichlorodiphenyltrichloroethane (DDT) prepared in liquid phase in KBr pellets and characterized using FT-IR and GC-MS followed by the application of DDT for molecular docking against human estrogen receptor alpha. The compound was modelled using GaussView software. Using Veda 04 program, the theoretical vibrational energy distributions and experimental vibrational frequencies were compared. Interestingly, C1 and C2�possess the highest atomic charge density distribution (ACDD)�of -0.284e and -0.283e while C21 and C11 have lowest ACDD of -0.064e and -0.063e in a relative manner, since the deactivating power of chlorine atoms decreases charge densities of the bonded carbon. The highest intramolecular interacting perturbation energy is 1121.92 kJ mol�1 occurs between ?*C19�C21 donor orbital and ?*C14�C16 acceptor orbital while the least intramolecular interaction occurs in the lone pair of LPC26 and the�sigma nonbonding (??C1�Cl24) NBO orbitals with E(2) of 32.21�kJ mol�1. Steric interaction was the only interaction found within the complex after the docking.

Experimental, DFT study, and in silico molecular docking investigations of dichlorodiphenyltrichloroethane against human estrogen receptor alpha

Advanced computational tools allowed to study a pure commercial sample of dichlorodiphenyltrichloroethane (DDT) prepared in liquid phase in KBr pellets and characterized using FT-IR and GC-MS followed by the application of DDT for molecular docking against human estrogen receptor alpha. The compound was modelled using GaussView software. Using Veda 04 program, the theoretical vibrational energy distributions and experimental vibrational frequencies were compared. Interestingly, C1 and C2�possess the highest atomic charge density distribution (ACDD)�of -0.284e and -0.283e while C21 and C11 have lowest ACDD of -0.064e and -0.063e in a relative manner, since the deactivating power of chlorine atoms decreases charge densities of the bonded carbon. The highest intramolecular interacting perturbation energy is 1121.92 kJ mol�1 occurs between ?*C19�C21 donor orbital and ?*C14�C16 acceptor orbital while the least intramolecular interaction occurs in the lone pair of LPC26 and the�sigma nonbonding (??C1�Cl24) NBO orbitals with E(2) of 32.21�kJ mol�1. Steric interaction was the only interaction found within the complex after the docking.