Molecules

The possible protective effects of methanolic extract Acrocarpus fraxinifolius leaves (MEAFL) were assessed against the APAP-induced organ toxicity in male rats. Also, the content of polyphenols extracted from AFL was studied, and their relationship with antioxidant activity was investigated. MEAFL was tested for cytotoxicity on Vero cell line, with reference to IC50, and other non-toxic concentrations of all the extracts. The antiviral activity against HSV1 for all non-toxic concentrations of the extract was determined using plaque reduction assay. It was found that MEAFL showed a reduction of serum hepatic and renal cellular toxicity and cellular lipid peroxidation, as well as enhanced cellular antioxidant. Also, our results revealed that the inhibitory activity of the virus was dose dependent on the polyphenol content of the examined extract. The MIC for the MEAFL extract was determined as well as the EC50 and SI. Calculated SI showed promising value for the MEAFL, and hence can be used as therapeutic medication for HSV1. To study other possible mode of action, Vero cells were treated with the examined extracts before, during, and after virus infection to give an insight on the interference of the extract in each step in the virus life cycle. In conclusion, MEAFL showed a remarkable antioxidant effect against APAP induced organs toxicity. Also, examined extracts exhibited the antiviral activity against HSV1.

NMR is a powerful tool for the structural and dynamic study of proteins. One of the necessary conditions for the study of these proteins is their isotopic labeling with 13C, 15N and sometimes 2H. One of the most widely used methods to obtain these labeled proteins is heterologous expression of the proteins in E. coli using 13C-D-glucose and 15NH4Cl as the sole nutrient source. In recent years, the price of 13C-D-glucose has almost tripled, making it essential to develop labelling methods that are as cost effective as possible. In this article, different parameters have been studied to achieve the most rational use of 13C-D-glucose and an optimised method has been developed to obtain labeled proteins with high labelling, low 13C-D-glucose consumption. Surprisingly, the optimised method is also simplier and does not require monitoring culture growth

The human voltage-gated sodium channel hNaV1.5 is essential for cardiac excitability. Though underrecognized, NaV1.5 is also expressed in multiple cancers, promoting cell migration and malignancy. hNaV1.5 is a therapeutic target but limited isoform specificity presents a risk of side effects via neuronal and skeletal muscle NaV channels. Here we identify Mg2+-protoporphyrin IX (MgPpIX), a Mg-containing tetrapyrrole and intermediate in chlorophyll biosynthesis, as inhibitor of hNaV1.5 (IC50 of 1 nM). The activity profile of various metal protoporphyrins correlates with the electrostatic potential at the metal center of the compounds. MgPpIX is specific to hNaV1.5, as no inhibition of other hNaV isoforms (hNaV1.2, 1.4, 1.7, 1.8) was detected. A mutagenesis study and structural modeling reveals that MgPpIX stabilizes the domain-II voltage sensor in the deactivated conformation, with residues E795 and N803 being relevant determinants. MgPpIX also inhibits native NaV channels in breast cancer MDA-MB-231 and colorectal carcinoma SW480 cell lines, and suppresses cell migration. MgPpIX is an exceptionally potent and specific inhibitor of hNaV1.5 and may serve as a lead compound in anti-cancer drug development.

The human DNA repair enzyme AlkB homologue-2 and 3 (ALKBH2 and ALKKBH3) repairs methyl adducts from genomic DNA. Overexpression of ALKBH2 and ALKBH3 has been implicated in both tumorigenesis and chemotherapy resistance in some cancers, including glioblastoma and renal cancer rendering it a potential therapeutic target and a diagnostic marker. However, no inhibitor is available against these important DNA repair proteins. Intending to repurpose a drug as an inhibitor of ALKBH2/ALKBH3, we performed in silico evaluation of HIV protease inhibitors and identified Ritonavir as an ALKBH2-interacting molecule. Using molecular dynamics simulation, we elucidated the molecular details of Ritonavir-ALKBH2 interaction. The present work highlights that Ritonavir might be used to target the ALKBH2-mediated DNA alkylation repair.

The renin-angiotensin system (RAS) comprises a biochemical cascade that hydrolyzes angiotensinogen into several different bioactive peptides, which can activate different receptors promoting plenty of specific effects. The aim of this study was to evaluate the presence of the putative product of alamandine, the pentapeptide alamandine-(1-5) in the circulation and its biological activity. To accomplish this we have used mass spectrometry (MALDI/TOF/TOF, LC-MS/MS) and several methodologies including isolated blood vessels, isolated perfused hearts, isolated cardiomyocytes, blood pressure recording in freely-moving normotensive and hypertensive rats (SHR), high resolution echocardiography (VEVO 2100), central administration (ICV infusion and microinjection in the insular cortex), cell culture (endothelial cells and GPCR-transfected CHO cells) and wild type and Mas, MrgD or AT2R deficient mice. Our results show that alamandine-(1-5) circulates in the human and rodent blood and promotes many biological central and peripheral actions. More importantly, its plasma concentration is increased in pediatric nephropathic patients. A major role for plasma ACE activity in the formation of alamandine-(1-5) from alamandine was observed using plasma samples from Angiotensinogen-KO mice. Alamandine-(1-5) increased Baroreflex sensitivity and produced a long-lasting ([~]6 hours) anti-hypertensive effect in SHR, associated with a significant reduction in cardiac output. A particularly important effect of this pentapeptide was observed in isolated perfused heart and cardiomyocyte contractility (reduced inotropism). It was capable of stimulating NO production through all receptors from the renin-angiotensin protective arm, (MAS, MrgD and AT2R) in CHO-transfected cells. Our data shows that Alamandine-(1-5) exhibits selective actions that set it apart from traditional concepts of the vasodilatory axis of the RAS and that are possibly intricately linked to a complex interplay between Mas, MrgD and AT2 receptors. This novel finding suggests that RAS may possess a complexity that surpasses our current understanding.

The receptor binding domain (RBD) of the spike protein of the Covid-19 virus is responsible for attachment to human ACE2. A number of recent articles have studied monoclonal antibody blocking [8-11] and peptide inhibitors [12-16] of the Covid-19 virus. Here we report virtual ligand-based screening that targets pockets on each side of an important binding site with residues 502-504 on the RBD that contact residues 353-357 [15] of hACE2. These ligands are intended as pre-exposure therapy for Covid-19 infection.

Diabetes mellitus is one of the complex metabolic disorders associated with individuals leading sedentary lifestyles. It leads to several complications rendering the normal function of vital organs like heart, liver, kidney, eye and brain. Scientists and doctors across the globe are involved in research for understanding the complex genetics of this disorder and formulating newer therapeutics accordingly. The finding of potential chemical entities and their underlying agonists or antagonist activities significantly controls the disorder but with some consequences. Thus there is demand for natural compounds and indigenous treatment methods for controlling the disorder with least or no adverse consequences. In the current work we present computational prediction of gamma-oryzanol as potential agonist of human peroxisome proliferator-activated receptor gamma (PPAR-{gamma}). A group of four gamma-oryzanol compound structures reported in PubChem database were downloaded and docked in the ligand binding site of five different human PPAR-{gamma} structures reported in PDB database. It was observed that most of the gamma-oryzanol compounds occupied themselves in the ligand binding P1, P2, P3, P4 sites with similar orientations as that of co-crystal agonists. Their binding conformations were assisted by some reasonable docking scores (-7 to -11 kcal/mol) and hydrogen bond interactions with some important conserved amino acid residues lining the ligand binding site. Additionally we have done a comparative molecular dynamics studies to reveal the flexibility of gamma-oryzanol in the ligand binding site in comparison to the co-crystal agonist and a scaffold analysis using the structure of six agonists and gamma-oryzanol for fetching potential scaffolds which may helpful in designing of new chemical entities.

Three copper(II) complexes containing 1,10-phenanthroline ([CuCl2(phen)]{middle dot}4H2O,1), neocuproine ([CuCl2(neo)]{middle dot}4H2O, 2) and tetramethyl-phenanthroline ([CuCl2(tmp)]{middle dot}4H2O, 3) as the primary ligand and another three copper(II) complexes with the L-Ala-Phe dipeptide as an auxiliary ligand: [Cu(L-Ala-Phe)(phen)]{middle dot}4H2O (4), [Cu(L-Ala-Phe)(neo)]{middle dot}4H2O (5) and [Cu(L-Ala-Phe)(tmp)]{middle dot}4H2O (6), inhibited cell viability in breast cancer MCF-7 cell line, both in the monolayer and spheroid cell culture models. The pair with tetramethyl-phenanthroline displayed a better selectivity index than cisPt and non-cytotoxicity-related ROS induction and apoptosis in the monolayer breast cancer model. Cell proliferation was affected by all compounds in a concentration-dependent manner, with a more substantial effect on the tetramethyl-phenanthroline complexes. Cell viability on multicellular spheroids showed a concentration-dependent reduction from 1 M, with IC50 that were half the one for cisplatin. All copper complexes, except for 1 showed DNA damage, demonstrated by the comet assay at a concentration below the IC50. The role of NHE1 has been linked to many types of cancers. Our study revealed that all compounds inhibited NHE1 activity in MCF-7 cells. However, only complexes containing the dipeptide auxiliary ligand could extend their effect on cell migration (Wound Healing Assay) and MMP-9 activity studied by zimography. Wester Blot analysis showed that expressions of MMP-2, MMP-9, and NHE1 were affected when MCF7 cells were treated with the six compounds as well. Overall, our results reveal an antitumor effect of all copper(II) complexes studied in breast cancer cells and a fundamental role of NHE1 in cell migration.

For the first time, we investigated through molecular docking analysis with Autodock Vina and Autodock 4 the Polydatin, a derivative of resveratrol, with Sodium/glucose cotransporter 2 (SGLT1) and Sodium/glucose cotransporter 2(SGLT2) and with Sirtuins proteins, reporting excellent results both in terms of binding energies scores and inhibition constant Ki. In particular, from our analyses, Polydatin appears to have an excellent energetic affinity with human SGLT2 on the one hand, and with the human Sirtuin6, even though, comparing the binding energy values with all the investigated proteins, no significant differences were found in termins of binding energies scores. An important aspect that we want to underline is that our computational analyzes, although very accurate, require investigations in Vitro, in Vivo, and clinical studies to confirm that Polydatin has a key role with SGLT2, SGLT1, and with the Sirtuin family.

A novel concept titled Peptide Mold for mapping potential binding sites in protein targets is presented. A large multiconformer tetrapeptide library comprising of 32 million conformations of all possible combinations of naturally-occurring amino acids was constructed and used for molecular docking analysis in the substrate-binding site of SARS-CoV-2 PLpro enzyme. The top-ranking, structurally-diverse tetrapeptide docked conformations (symbolizing peptide mold, analogous to a clay mold) were used then for elucidating a five-point pharmacophore. Ligand-based virtual screening of a large, multiconformer library of phytoconstituents using the derived five-point pharmacophore led to identification of potential binders for SARS-CoV-2 PLpro at its substrate-binding site. The approach is based on generating the imprint of a macromolecular binding site (cavity) using tetrapeptides (clay), thereby generating a reverse mold (with definitive shape and size), which can further be used for identifying small-molecule ligands matching the captured features of the target binding site. The approach is based on the fact that the individual amino acids in the tetrapeptide represent all possible drug-receptor interaction features (electrostatic, H-bonding, van der Waals, dispersion and hydrophobic among others). The peptide mold approach can be extended to any protein target for mapping the binding site(s), and further use of the generated pharmacophore model for virtual screening of potential binders. The peptide mold approach is a robust, hybrid computational screening strategy, overcoming the present limitations of structure-based methods, e.g., molecular docking and the ligand-based methods such as pharmacophore search. Exploration of the peptide mold strategy is expected to yield high-quality, reliable and interesting virtual hits in the computational screening campaigns during the hit and lead identification stages.

Head and neck squamous cell carcinoma (HNSCC) is a therapeutically challenging cancer what underscoring the need for new chemical agents that selectively induce programmed cell death. Fisetin, a naturally occurring flavonoid, exhibits promising anticancer activity but displays limited proapoptotic efficacy and selectivity. Here, we examined whether alkoxylated modification of fisetin enhances its ability to induce apoptosis in HNSCC cells. Fisetin derivatives bearing four-carbon substituents were synthesized and evaluated in multiple HNSCC cell lines. Two derivatives, MKT218 and MKT257, markedly reduced HNSCC cell viability at low micromolar concentrations with low toxicity towards normal human fibroblasts. Notably, the observed cytotoxicity was not associated with activation of a canonical DNA damage response, as neither {gamma}H2AX accumulation nor p53 activation was detected. Furthermore, PARP1 cleavage and live-cell imaging combined with annexin V/EthD-III staining revealed a significantly higher proportion of apoptotic cells. The effect was stronger following treatment with MKT218 and MKT257 compared with fisetin. Time-lapse microscopy further demonstrated that fisetin derivatives, particularly MKT218, promote mitosis-associated apoptosis, in contrast to the predominantly cytostatic effect of fisetin. Moreover, in silico docking suggested that MKT218 exerts its pro-apoptotic activity through a multi-target interaction profile involving key regulators of cell survival and apoptosis rather than a single dominant target. To sum up, our findings suggest that alkoxylated fisetin derivatives may be constituted as new non-genotoxic inducers of apoptosis in HNSCC cells.

1Members of the parvalbumin (PV) family of calcium binding proteins are found in a variety of vertebrates, where can they influence neural functions, muscle contraction and immune responses. It was reported that the -parvalbumin (PV)s AB domain comprising two -helices, dramatically increases the proteins calcium (Ca2+) affinity by {approx}10 kcal/mol. To understand the structural basis of this effect, we conducted all-atom molecular dynamics (MD) simulations of WT PV and truncated -parvalbumin ({Delta}PV) constructs. Additionally, we also examined the binding of magnesium (Mg2+) to these isoforms, which is much weaker than Ca2+ (Mg2+ actually does not bind to the {Delta}PV). Our key finding is that reorganization energies (RE) assessed using molecular mechanics generalized Born approximation (MM/GBSA) correctly rank-order the variants according to their published Ca2+ and Mg2+ affinities. The [Formula] of the {Delta}PV compared to the wild-type (WT) is 415.57{+/-}0.55 kcal/mol, indicating that forming a holo state of {Delta}PV in the presence of Ca2+ incurs a greater reorganization penalty than the WT. This is consistent with the {Delta}PV exhibiting lesser Ca2+ affinity than the WT ({approx}9.5 kcal/mol). Similar trend was observed for Mg2+ bound variants as well. Further, we screened for metrics such as oxygen coordination of EF hand residues with ions and found that the total oxygen coordination number (16 vs. 12 in WT:Ca2+ and {Delta}PV:Ca2+) correlate with the reported ion affinities (-22 vs. -12.6 kcal/mol in WT:Ca2+ and {Delta}PV:Ca2+), which indicates that AB domain is required for the protein to coordinate with maximal efficiency with the binding ions. To our surprise, no significant differences were observed between the Mg2+ bound WT and {Delta}PV isoforms. Additionally, we have screened for factors such as total number of waters, hydrogen bonds, protein helicity and {beta}-content for the entire protein, which enables us to understand the impact of lack of AB domain on the entire structure and not just binding sites. Our data indicate that AB improves the overall helicity ({approx}5%) in apo as well as holo forms. Particularly, AB increases -helicity in the D-helix residues (i.e., 60-65) upon ion binding by {approx}35% (90% vs. 55% in the Ca2+ bound WT and {Delta}PV, 60% vs. 20% in the Mg2+ bound WT and {Delta}PV), which likely contributes to high Ca2+ binding affinity. On the contrary, no significant effect on the overall {beta}-content was observed. Similarly, increased dehydration ({approx}50) and increase in total number of hydrogen bonds ({approx}7) were observed upon ion binding in both the WT and {Delta}PV systems, however, no significant differences were observed between the WT and {Delta}PV variants and also between Ca2+ and Mg2+ isoforms. We speculate that this is due to the partially folded apo state that was captured in our MD simulations, which might not be physiologically relevant as suggested by NMR experiments [1]. Also, we have identified seven different interactions that might play a key role in binding the AB domain with the CDEF helices, particularly the D22(AB)-S78(CDEF) hydrogen bond. Overall, this study indicates that local (i.e., the EF hands) as well as global factors play a role in improved ion binding due to AB domain.

Curcumin and trans-cinnamaldehyde are acrolein-based Michael acceptor compounds that are commonly found in domestic condiments, and known to cause cancer cell death via redox mechanisms. Based on the structural features of these compounds we designed and synthesized several 2-cinnamamido-N-substituted-cinnamamide (bis-cinnamamide) compounds. One of the derivatives, (Z)-2-[(E)-cinnamamido]-3-phenyl-N-propylacrylamide (1512) showed a moderate antiproliferative potency (HT116 cell line inhibition of 32.0 {micro}M {+/-} 2.6) with proven cellular activities leading to apoptosis. Importantly, 1512 exhibited good selectivity toxicity on cancer cells over noncancerous cells (IC50 of C-166 cell lines >100 {micro}M), and low cancer cell resistance at 100 {micro}M dose (growth rate 10.1{+/-}1.1%). We subsequently carried out structure activity relationship studies with 1512. Derivatives with electron rich moiety at the aryl ring of the 2-aminocinnamaide moiety exhibited strong antiproliferative action while electron withdrawing groups caused loss of activity. Our most promising compound, 4112 [(Z)-3-(1H-indol-3-yl)-N-propyl-2-[(E)-3-(thien-2-yl)propenamido)propenamide] killed cancer cells at IC50 = 0.89 {+/-} 0.04 {micro}M (Caco-2), 2.85 {+/-} 1.5 (HCT-116) and 1.65 {+/-} 0.07 (HT-29), while exhibiting much weaker potency on C-166 and BHK normal cell lines (IC50 = 71 {+/-} 5.12 and 77.6 {+/-} 6.2 {micro}M, respectively). Cellular studies towards identifying the compounds mechanism of cytotoxic activities revealed that apoptotic induction occurs in part due to oxidative stress. Importantly, the compounds showed inhibition of cancer stem cells that are critical for maintaining the potential for self-renewal and stemness. The results presented here show discovery of Michael addition compounds that potently kill cancer cells by a defined mechanism, with minimal effect on normal noncancerous cell.

AbstractionCurcumin (CUR), a naturally occurring phenolic small molecule, has been extensively applied in the treatment of diverse diseases for over half a century. Currently, this chemical has also been verified to exhibit a wide array of its biopharmacological activities, such as enhancing immunity, possessing antiviral, anti-cardiovascular, and anti-cancer properties. Besides, also as a nutritional supplement, CUR enables inflammatory prevention and enhance the combined efficacy of it with chemotherapy. Such characteristic of safeguarding normal organisms while treating or alleviating diseases is manifested as a common bidirectional regulation and its hormetin effect of phenolic compounds, particularly in response to stress. In this redox process, two antioxidant transcription factors Nrf1 and Nrf2 (encoded by Nfe2l1 and Nfe2l2, respectively) play distinct and crucial roles. Thereby, we investigated their expression profiles of genes regulated by Nrf1 and Nrf2 in different signaling responses modulated by CUR. The resulting evidence has been provided, demonstrating that distinctive cellular metabolisms, molecular pathways, and signaling mechanisms account for Nrf1 and Nrf2, as drug targets. Both factors also play diverse roles in the anticancer effects of CUR on HepG2 cells and xenograft mice. However, the effect of CUR on xenograft tumors in vivo is not entirely satisfactory, although such anti-cancer effect was achieved by promoting Nrf1 expression, it appeared more reliant on Nrf2 (particularly in the absence of Nrf1). As such, we unexpectedly discovered there is not a simple regulatory relationship between CUR and Nrf2. This is supported by substantial inhibition of Nrf2 by CUR in the aberrantly proliferating Nrf1-/- cells, even albeit this chemical stimulates the expression of Nrf1 and Nrf2 in wild-type cells.

This manuscript presents an updated version of PEPlife, a manually curated database that provides comprehensive information on peptide half-life. The updated version, PEPlife2, includes 4,412 entries, compared to 2,229 entries in the previous version. Each entry contains detailed information that include experimental methods used to determine half-life, chemical modifications, biological activity, routes of administration, and other relevant data. The database encompasses 402 proteins and 1,781 peptides, derived from 105 unique assays. In addition to natural peptide sequences, PEPlife2 includes cyclic peptides and chemically modified peptides, such as those with N- and C-terminal modifications. To provide structural insights, peptide and protein structures are sourced from the Protein Data Bank (PDB) or predicted using PEPstrMOD. To support the scientific community, we have developed a user-friendly interface and integrated advanced analytical tools, including BLAST, Smith-Waterman, GGSEARCH, CLUSTALW, and MUSTANG. The updated database is accessible at: https://webs.iiitd.edu.in/raghava/peplife2/. Authors BiographyO_LIUrooj Alam is currently pursuing a Masters degree in Computational Biology at the Department of Computational Biology, Indraprastha Institute of Information Technology, New Delhi, India. C_LIO_LIKunal Chaudhary is working as Ph.D. in Computational biology from Department of Computational Biology, Indraprastha Institute of Information Technology, New Delhi, India C_LIO_LINishant Kumar is currently working as Ph.D. in Computational biology from Department of Computational Biology, Indraprastha Institute of Information Technology, New Delhi, India C_LIO_LIRitu Tomer is currently working as Ph.D. in Computational biology from Department of Computational Biology, Indraprastha Institute of Information Technology, New Delhi, India C_LIO_LISumeet Patiyal is currently working as a postdoctoral visiting fellow Cancer Data Science Laboratory, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA. C_LIO_LIGajendra P. S. Raghava is currently working as Professor and Head of Department of Computational Biology, Indraprastha Institute of Information Technology, New Delhi, India. C_LI

Hylocereus polyrhizus, locally known as red dragon fruit, is valued for its nutritional benefits including high levels of antioxidants and is gaining popularity as both a food and a medicinal plant. The present study addressed the in vivo and in silico chemopreventive potential of Hylocereus polyrhizus peel extract (HPPE) in DMBA-croton oil induced skin carcinogenic model mice. The mRNA expression level of pro-inflammatory cytokines and inflammatory mediators in tumor mass were estimated by real time-RT-qPCR. In addition, molecular docking and molecular dynamic simulation analyses were conducted on the reported compounds. In the in vivo chemopreventive activity assessment, the peel extract at 500 mg/kg was found effective in reducing total tumor number, yield, burden, incidence, and weight. Total proteins and endogenous antioxidants (GSH, SOD, CAT) levels in liver and skin tissues from mice were significantly (P<0.05) elevated. In addition, the HPPE at 500 mg/kg dose significantly reduced (P<0.05) the gene expression of pro-inflammatory cytokines such as TNF-, IL-1{beta}, IL-6, IL-18, and inflammatory mediators like TGF-{beta}1, COX-2, and NF{kappa}B. In the molecular docking studies, reported compounds including Quercimeritrin, Rutin, and Kaempferol 3-O-{beta}-D-glucopyranoside were identified as the top-performing compounds, with a docking score of - 7.4, -7.1 and -7.0 kcal/mol against TGF-{beta}1 protein. This indicates stronger binding interactions compared to vincristine (-5.3 kcal/mol). In drug-likeness assessment, all compounds demonstrated the most favourable ADMET and pharmacokinetic profile. Furthermore, MDS data showed greater dynamic stability for Kaempferol 3-O-{beta}-D-glucopyranoside and Rutin, while vincristine exhibited higher fluctuations. The results suggest that HPPE may serve as a potential inhibitor of skin carcinogenesis through upregulating endogenous antioxidants as well as suppressing different proinflammatory and inflammatory cytokines. Quercimeritrin, Rutin, and Kaempferol 3-O-{beta}-D-glucopyranoside might be the probable leads responsible for this chemopreventive activity. Highlights{checkmark} Hylocereus polyrhizus peel extract (HPPE) demonstrated notable chemopreventive potential against skin cancer. {checkmark}HPPE significantly increased endogenous antioxidants (GSH, SOD, CAT) in liver and skin tissues, suggesting enhanced cellular defense mechanisms. {checkmark}Gene expression of pro-inflammatory cytokines (TNF-, IL-1{beta}, IL-6, IL-18) and inflammatory mediators (TGF-{beta}1, COX-2, NF{kappa}B) were notably suppressed by HPPE. {checkmark}Quercimeritrin (CID: 5282160) demonstrated strong binding to TGF-{beta}1 with a docking score of -7.4 kcal/mol, outperforming vincristine (-5.3 kcal/mol). {checkmark}Kaempferol 3-O-beta-D-glucopyranoside (CID: 5282102) and rutin (CID: 5280805) showed the best ADMET, pharmacokinetic profiles, and molecular stability, supporting their potential as lead compounds.

Breast cancer (BC) is a prevalent form of cancer observed in women across the globe, constituting over a quarter of all female BC cases. The treatment of BC continues to require significant efficacy, aiming to achieve high success rates while minimizing adverse effects on the body as a whole. In the current study, 3-epicaryoptin was tested for the molecular mechanism of its anti-cancer activity in the human breast cancer cell line, MCF-7. We investigated cell viability by MTT assay, cell cycle kinetics and apoptosis, immunofluorescence straining, molecular modelling, and ADMET profiling. MTT assay results showed that 3-epicaryoptin was found cytotoxic against MCF-7 cells with an IC50 value of 344.64 {micro}g mL-1 for 48 h. Flow cytometric analysis exhibited that 3-epicaryoptin halted the MCF-7 cells in the G2/M phase and subsequently induced apoptosis in a time-dependent manner. Our immunofluorescence studies indicated that 3-epicaryoptin inhibited microtubule polymerization in MCF-7 cells. Furthermore, molecular docking followed by molecular dynamics (MD) simulation studies demonstrated the ability of 3-epicaryoptin to interact with the tubulin protein at the colchicine binding pockets. Overall, our results suggest that 3-epicaryoptin can inhibit the proliferation of human breast cancer cells by depolymerizing of cellular microtubule networks, which causes cell cycle arrest and promotes apoptotic cell death. Therefore, it has been indicated that the natural product 3-epicaryoptin exhibited considerable promise as a potent therapeutic agent capable of inducing apoptosis in breast cancer cells.

p53 protein coded by the Tp53 gene is considered as one of the most intensively researched protein and mainly due to its role as a tumor suppressor, it acts as a tumor suppressor by carrying out two biologically complex processes namely Cell cycle arrest and apoptosis, In the oncogenic Y220C mutant p53, tyrosine is replaced by cysteine at 220th residue of the DNA binding Domain which causes the formation of a surface crevice, this specific mutation is responsible for approx. 100,000 cancer cases per year due to the destabilization and denaturation of the protein, as a result, the protein degrades at room temperature. In this work we carry out intensive Molecular Dynamic Simulations and Molecular Docking Studies to understand the structural dynamics of wild type p53 and changes the occurs in the mutant protein and also try to design lead against the druggable crevice and at the end of our study we used fragment-based optimization to come up with lead molecules which can act as scaffold for further drug development process

Targeting apoptosis is a promising approach to inhibit the abnormal cell proliferation of cancer progression. Existing anti-apoptotic drugs, many derived from chemical substances, have often failed to combat cancer development and progression. Therefore, identification of apoptosis-inducing anticancer agents from plant-derived sources has become a key aim in cancer research. The present study was designed to explore the regulation of apoptosis by Tabebuia pallida (T. pallida) using an Ehrlich Ascites Carcinoma (EAC) mouse model and compositional analysis by LC-ESI-MS/MS. Dried and powdered T. pallida leaves (TPL), stem bark (TPSB), root bark (TPRB) and flowers (TPF) were extracted with 80% methanol. Using cultured EAC cells and EAC-bearing mice with and without these extracts, anticancer activities were studied by assessing cytotoxicity and tumor cell growth inhibition, changes in life span of mice, and hematological and biochemical parameters. Apoptosis was analyzed by microscopy and expression of selected apoptosis-related genes (Bcl-2, Bcl-xL, NF{kappa}-B, PARP-1, p53, Bax, caspase-3 and -8) using RT-PCR. LC-ESI-MS analysis was performed to identify the major compounds from the most active extracts. In EAC mice compared with untreated controls, the TPL extract exhibited the highest cytotoxicity with significant tumor cell growth inhibition (p< 0.001), reduced ascites by body weight (p< 0.01), increased the life span (p<0.001), normalized blood parameters (RBC/WBC counts), and increased the levels of superoxide dismutase and catalase. TPL-treated EAC cells showed apoptotic characteristics of membrane blebbing, chromatin condensation and nuclear fragmentation, and caspase-3 activation, compared with untreated EAC cells. Moreover, annexin V-FITC and propidium iodide signals were greatly enhanced in response to TPL treatment, indicating apoptosis induction. Pro- and anti-apoptotic signaling after TPL treatment demonstrated up-regulated p53, Bax and PARP-1, and down-regulated NF{kappa}-B, Bcl-2 and Bcl-xL expression, suggesting that TPL shifts the balance of pro- and anti-apoptotic genes towards cell death. LC-ESI-MS data of TPL showed a mixture of glycosides, lapachol, and quercetin antioxidant and its derivatives that were significantly linked to cancer cell targets. In conclusion, the TPL extract of T. pallida possesses significant anticancer activity. The tumor suppressive mechanism is due to apoptosis induced by activation of antioxidant enzymes and caspases and mediated by a change in the balance of pro- and anti-apoptotic genes that promotes cell death. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=150 SRC="FIGDIR/small/426226v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@1112faaorg.highwire.dtl.DTLVardef@1505d1dorg.highwire.dtl.DTLVardef@1115a2aorg.highwire.dtl.DTLVardef@36a915_HPS_FORMAT_FIGEXP M_FIG C_FIG

One of the primary health issues caused by inadequate blood sugar regulation is Diabetes Mellitus (DM). Diabetes and its consequences remain clinically significant even with the development of oral hypoglycemic medications. Therefore, Ficus racemosa (F. racemosa) plant has been studied for assessing of its antidiabetic potential coupling with animal model and in silico experiments. Drug Alloxan (150 mg/kg) was injected to induce the experimental diabetes in Swiss Albino mice, and two doses methanol extract of the F. racemosa fruit (300 and 500 mg/kg) along with glibenclamide (5 mg/kg) were given orally. Oral Glucose Tolerance Test (OGTT) and acute toxicity were performed as well in both diabetic and non-diabetic mice. Later, in silico experiments including ADMET profiling, molecular docking and simulations were performed. The administration of a dosage less than 3000 mg/kg has been observed to be well-tolerated by mice, with no reported instances of mortality or adverse effects. Following oral administration for 7 days, the blood glucose level (BGL) was significantly decreased in mice model in both doses of extracts, indicating the effect of F. racemosa. Subsequent to this, molecular docking and simulations have indicated that the SIRT1 receptor exhibits a higher binding affinity towards four specific compounds, namely Friedelin, Lupeol Acetate, Gluanol, and Ferulic Acid, as indicated by the dynamics parameters and interacting residues. The current investigation provided evidence that the fruit extract of F. racemosa significantly mitigated the hyperglycemic impact. Moreover, a total of four substances have been found that play a crucial role in the mechanisms behind the reduction of diabetic effects. Hence, the current investigation could potentially serve as a viable therapeutic approach in the treatment of diabetes.