Greater Binding Affinity Research Articles

Protective Effect of Quercetin and p-Coumaric Acid (p-CA) Against Cardiotoxicity: An In Silico Study.

Hydroxychloroquine (HCQ) is a common antimalarial drug that has been used effectively in the treatment of various rheumatic and auto-immunity diseases. The major side effects and drawbacks associated with HCQ are cardiotoxicity, retinopathy, gastrointestinal upset, and neuromyopathy however, cardiotoxicity is an increasing concern and it is critical to avoid heart dysfunction induced by HCQ. The present work is focused on receptor and signaling molecules associated with pathways attributing to drug-induced cardiotoxicity. We analyzed the therapeutic efficacy of selected natural products in HCQ-induced cardiotoxicity through insilico. We selected Syzygium cumini polyphenols, quercetin, and p-coumaric acid. The motivation behind selecting quercetin, and p-coumaric acid is their wide applicability as an antioxidative, anti-inflammatory, antiapoptotic, and cardioprotective. For predicting quercetin, p-coumaric acid, and HCQ toxicity and physicochemical properties, in silico studies were performed using ProTox II and Swiss ADME. We further performed molecular docking using Autodock Vina and Discovery Studio visualizer to find the affinity of selected polyphenols against signaling molecules and receptors. Then we performed network pharmacological studies of selected signaling molecules. We analyzed that the computational method indicated quercetin (Δ G -9.3 kcal/mol) has greater binding affinity than p-Coumaric acid for prevention and restoration of the disease while hydroxychloroquine was taken as a control. It can be concluded that Syzygium cumini, polyphenols may aid in the future therapeutic potential against HCQ-induced cardiotoxicity.

In Vitro and in Silico Analysis of Phytochemicals from Asteraceae Plant Species Against Wound-Infecting Pathogens

The idea of polyherbal combinations has a strong foundation in modern medicine and has had amazing success, givingpatients new hope. Polyherbal mixtures have managed various conditions, from cancer to autoimmune disorders. Given that it hasalready been shown to be secure and efficient, this therapeutic approach is gaining popularity. In alternative, complementarytherapies, polyherbal formulations blend many herbs to provide an additive effect. Physicians are accepting this strategy, which hasbeen used for generations. Numerous research papers in the pharmaceutical field demonstrate the efficacy of combining plants andthe present study aimed to evaluate antibacterial phytochemicals from a polyherbal extract. Phytoconstituents extracted with ethanoland ethyl acetate by conventional soxhlet method and chemical constituents were analyzed by qualitative and GC-MS. Antibacterialactivity of extracts tested by disc diffusion method against wound infecting pathogens. MIC was done by Alamar blue assay. Inhibitionof ESBL enzyme predicted by in silico docking using Autodock. Extraction with ethanol shows less significant positive results thanethylacetate in qualitative and antibacterial studies. GC-MS analysis showed the presence of 35 bioactive compounds from ethylacetate and 45 compounds from ethanol. Cyclononasiloxane, Octodecane, hexadecanoic acid, and Benzenedicarboxylic werecommonly found in both extracts. The ethylacetate PHE exhibited strong antibacterial activity and was more effective than thestandard. From GCMS, three compounds were predicted to have good ADME characteristics with high gastrointestinal absorptionand metabolic activity. Compound 6-(3-fluorobenzylamino) purine has been selected and investigated by in silico method, showingthat it has a greater binding affinity with ESBL, ompC, and murA after being docked. The ethyl acetate extract demonstrated goodaction against certain pathogenic bacteria in the current experiment, while the ethanol extract exhibited no activity. Our findingsconclude that the polyherbal extracts have broad-spectrum activity against bacteria and depend on the solvent used.

Molecular mechanism of Yi-Qi-Yang-Yin-Ye against obesity in rats using network pharmacology, molecular docking, and molecular dynamics simulations

The increasing prevalence of obesity globally, which as well as affecting people's daily lives and increasing the risk of obesity complications, also threatens the health of animal organisms simultaneously. It’s been reported that Yi-Qi-Yang-Yin-Ye had remarkable efficacy in the treatment of obesity. The specific underlying mechanism of action of Yi-Qi-Yang-Yin-Ye in treating obesity, however, remains ambiguous. Therefore, the innovative approach, which is network pharmacology combined with molecular docking and molecular dynamics simulations, was employed in the current research to explore the potential mechanism and promote further development in the treatment of obesity. The active ingredients and related targets of Yi-Qi-Yang-Yin-Ye and related targets of obesity were summarized from extensive public databases. Furthermore, network topology analysis and pathway enrichment analysis were performed to explore the complicated interactions between drug and targets. Finally, accurate validation methods composed of molecular docking and molecular dynamics simulations were conducted to elucidate the binding affinity of Yi-Qi-Yang-Yin-Ye with obesity-related targets. As a result, 13 main active ingredients and 5 core targets of Yi-Qi-Yang-Yin-Ye against obesity in rats were acquired through primary screening of network topology analysis. Pathway enrichment analysis demonstrated that intersectional targets were involved in multiple signaling pathways, where PI3K-Akt signaling pathway, MAPK signaling pathway, and Insulin resistance were the main pathways of Yi-Qi-Yang-Yin-Ye in treating obesity in rats. Finally, molecular docking indicated that the seven critical active ingredients displayed great binding affinity to the hub targets. Furthermore, molecular dynamics simulations further screened and obtained that five critical active ingredients acting on the Mapk1 target for Yi-Qi-Yang-Yin-Ye against obesity in rats. The innovative approach and the results achieved have further contributed to and revealed the molecular mechanisms for treating obesity, providing an alternative for treating obesity in animals and humans.

Cardioprotective effect of tetra(aniline) containing terpolymers through miR-15a-5p and MFN-2 regulation against hypertrophic responses

ObjectiveCardiac hypertrophy is a condition of abnormal cardiomyocyte enlargement accompanied by ventricular wall thickening. The study aims to investigate the role of miR-15a-5p in the regulation of mitofusin-2 (MFN-2) and to explore the cardioprotective effect of terpolymers ES-37 and L-37. MethodsIn this study, the Sprague Dawley rats’ cardiac hypertrophic model was established by administering 5 mg/kg Isoproterenol subcutaneously every other day for 14 days. As treatment rats received NAC (50 mg/kg), NAC treatment (50 mg/kg NAC + 5 mg/kg ISO), ES-37 (1 mg/kg) and ES-37 treatment (1 mg/kg ES-37+5 mg/kg ISO), L-37 (1 mg/kg) and L-37 treatment (1 mg/kg L-37+5 mg/kg ISO). subcutaneously every other day for 14 days. NAC, ES 37 and L-37 were given after 1 h of Isoproterenol administration in treatment groups. Cardiac hypertrophy was confirmed through morphological and histological analysis. For estimation of oxidative stress profiling, ROS and TBARS and antioxidative profiling superoxide dismutase (SOD), Catalase, and Glutathione (GSH) levels were checked. Triglyceride, cholesterol, alanine transaminase (ALT), and aspartate transaminase (AST) were performed to evaluate levels of lipid profiling and liver profiling. Molecular expression analysis was checked through real-time PCR, and western blotting both at the transcriptional and translational levels. Molecular docking studies were performed to study the interactions and modes of binding between the synthetic polymers with three proteins (Mitofusin-2, DRP-1 and PUMA). All the studies were carried out using the AutoDock Vina software and the protein-ligand complexes were visualized in Biovia Discovery Studio.Cardiac hypertrophy was confirmed by the relative changes in the cellular structure of the heart by histopathological examination and physiological changes by estimating organ weights. Biochemical profiling results depict elevated oxidative and lipid profiles signify myocardial damage. N-acetyl cysteine (NAC), ES-37, and L-37 overcome the cardiac hypertrophic responses through attenuating oxidative stress and enhancing the antioxidative signaling mechanism. miR-15a-5p was identified as hypertrophic microRNA directly regulating the expression of Mitofusin-2 (MFN-2). Significantly increased expression of miR-15a-5p, Dynamin related protein 1 (Drp1), and P53 upregulated modulator of apoptosis (PUMA), was observed in the disease group, whereas MFN-2 expression was observed downregulated. N-acetyl cysteine (NAC), ES-37, and L-37 showed increased expression of antiapoptotic maker MFN-2 and decreased expression of miR-15a-5p, Drp1, and PUMA in treatment groups suggesting their cardioprotective role in attenuation of cardiac hypertrophy. An analysis of the docking results shows that ES-37 has greater binding affinity with the target proteins compared to L-37, with the highest binding values reported for MFN-2. ConclusionThe physiochemical properties of ES-37 and L-37 predicted it as a good drug-like molecule and its mechanism of action is predictably through inhibition of ROS. Molecular docking results shows that the polymer ES-37 has greater binding affinity with the target proteins compared to L-37, with the highest binding values reported for MFN-2. Thus, the study validates the role and targeting of miR-15a-5p and MFN-2 in cardiac hypertrophy as well as the therapeutic potential of NAC, ES-37, and L-37 in overcoming oxidative stress and myocardial damage.

Synthesis, Characterization, Biological Evaluation, DFT Studies and Molecular Docking of Novel 12-(2-(1H-Imidazol-1-yl)quinolin-3-yl)-8-methyl-2,3,4,12-tetrahydro- 1H-benzo[4,5]thiazolo[2,3-b]quinazolin-1-one and its Derivatives

Twelve new compounds were synthesized consisting of biquinoline-imidazole-benzothiazole hybrids 6a-l using a base catalyzed one pot MCR reaction, these were then screened for in vitro biological activities against cancer cell lines. Among the twelve prepared potential anticancer agents, compound 6j was found to be the most active against EGFR (IC50 of 0.14 ± 0.03 Μm), A549 and HepG2, also the same derivative of the series was found to be most active against FabH (IC50 of 3.1 Μm) E. coli. Further the molecules were analysed for DFT and molecular docking studies to calculate the distance and angle between the active parts of the molecules as well as charge density distribution over the molecules which provides a qualitative measure for the effective the binding of molecules in the active pockets with greater binding affinity.

Simultaneous binding characterization of different chromium speciation to serum albumin.

The binding process between three species of chromium and serum albumin (SA) was investigated, as well as the interaction between K2Cr2O7 and bovine serum albumin (BSA) under coexistence of different chromium forms. CrCl3, K2Cr2O7 and Crpic bound to SA spontaneously through Van der Waals force, and their binding constants were 103-104M-1 at 298K, respectively. K2Cr2O7 and Crpic both had strong binding affinity for BSA, and significantly affected the secondary structure of BSA and the microenvironment surrounding amino acid residues. Chromium exhibited a greater fluorescence quenching constant towards HSA than toward BSA, and K2Cr2O7 induced greater conformational changes in human serum albumin (HSA) than in BSA. A weak binding of CrCl3 to BSA had no significant effect on the binding affinity of K2Cr2O7 to BSA. K2Cr2O7 and BSA have a greater binding affinity when coexisting with Crpic, and K2Cr2O7 induces a greater conformational change in BSA.

Anticaries Potential of Temu Kunci-Serai Ethyl Acetate Extract Combination: In Vitro and Molecular Studies Approach

Dental caries is one of the problems in dental disorders suffered by many people. There have been many ways of handling such as using temu kunci and lemongrass plants. This study aims to determine the antibacterial activity of caries in vitro and predict the mechanism of action of the bacteria that cause caries Streptococcus mutans. Temu kunci and lemongrass were extracted using ethyl acetate solvent. antibacterial tests against Streptococcus mutans were carried out using diffusion tests with a combination treatment of temu kunci: serai extracts 5% b/v: 5% b/v, 5% b/v: 10% b/v, and 10% b/v: 5% b/v. The positive control used amoxicillin and DMSO as a negative control. Potency as an anticaries drug of drived compound from Temu Kunci and Serai were evaluated by molecular docking using glucosyltransferase (3AIC). The results showed that the combination of temu kunci and serai with concentration 10% b/v: 5% b/v has potential as anticaries against Streptococcus mutans. Molecular studies depicted that Panduratin A, Isopanduratin, and 1,3-O-di-p-coumaroylglycerol have great activity toward 3AIC, respectively. Especially for Panduratin A and Isopanduratin, those compound depicted great and similar binding affinity (-8.4 kcal mol-1) that lower than Acarbose as native ligand (-8.3 kcal mol-1). Furthermore, those compound binding similarity ilustrated activity mechanism similarly with native ligand toward receptor. Additionally, the profiling drug-target interaction suggested Temu Kunci’s derived compounds have great potential as anticaries treatment.

Inhibition of OAT1/3 and CMPF uptake attenuates myocardial ischemia-induced chronic heart failure via decreasing fatty acid oxidation and the therapeutic effects of ruscogenin

Chronic heart failure (CHF) as a long-term disease is highly prevalent in elder people worldwide. Early diagnosis and treatments are crucial for preventing the development of CHF. Herein, we aimed to identify novel diagnostic biomarker, therapeutic target and drug for CHF. Untargeted metabolomic analysis has been used to characterize the different metabolomic profile between CHF patients and healthy people. Meanwhile, the targeted metabolomic study demonstrated the elevation of 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF) in the serum of CHF patients and coronary artery ligation-induced CHF mice. Subsequently, we firstly observed that elevation of CMPF impaired cardiac function and aggravated myocardial injury by enhancing fatty acid oxidation (FAO). Interestingly, inhibition of responsible transporters organic anion transporter 1/3 (OAT1/3) has been found to decrease the CMPF level, and suppress FAO-related key protein expressions including peroxisome proliferator-activated receptor alpha, peroxisome proliferative activated receptor-α, carnitine palmitoyl transferase 1, and malonyl CoA decarboxylase in coronary artery ligation-induced CHF mice. Meanwhile, the inhibitor of OAT1/3 presented an excellent improvement in cardiac function and histological injury. Based on the above findings, molecular docking was adopted to screen the potential therapeutic drug targeting OAT1/3, and ruscogenin (RUS) exhibited a great binding affinity with OAT1 and OAT3. Next, it was verified that RUS could remarkedly decrease the expression of OAT1/3 and CMPF levels in heart tissue of CHF mice, as well as suppress the expression of FAO-related proteins. What's more, RUS can effectively improve cardiac function, myocardial fibrosis and morphological damage. Collectively, this study provided a potential metabolic marker CMPF and novel target OAT1/3 for CHF, which were demonstrated to be involved in FAO. And RUS was identified as a potential anti-FAO drug for CHF by regulating OAT1/3.

Synthesis, characterization and molecular docking study of pyrazolines synthesized from chalcones: Antioxidant and acetylcholinesterase activities

Pyrazolines are heterocyclic compounds with nitrogen-containing rings, known mainly for their pharmacological potential. In recent times, much attention has been paid to the synthesis of these compounds to discover a wide spectrum of biological activities. Therefore, thirteen pyrazoline derivatives were synthesized from a cyclization reaction between chalcones and phenylhydrazines and showed total yields of 56 - 90%. All compounds were fully characterized by Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (1H and 13C NMR), and gas chromatography/mass spectrometry (GC/MS). Compounds were submitted to antioxidant activity using DPPH assay and acetylcholinesterase tests. It was observed that different substituents on the A and B rings of the pyrazolines have an influence on the antioxidant activity. Compounds 3, 7, 11 and 13 showed good activity with IC50 0.4017, 0.2892, 0.1100 and 0.0578, respectively. In the acetylcholinesterase enzyme inhibition assay, compounds 8 and 9 showed very good inhibition, similar to the used standards. The synthesized compounds that were shown to be active in the in vitro assay of acetylcholinesterase inhibition were investigated and subjected to molecular docking studies, and the results obtained showed great binding affinity and the best pose for compounds 3-(4-bromophenyl)-1,5-diphenyl-4,5-dihydro-1H-pyrazole (6) and 3-(4-chlorophenyl)-5-(4-methoxyphenyl)-1-phenyl-4,5-dihydro-1H-pyrazole (8), according to the results experimental.

Inhibitory potential of phytochemicals on five SARS-CoV-2 proteins: in silico evaluation of endemic plants of Bosnia and Herzegovina

The outbreak of the coronavirus disease 2019, caused by the SARS-CoV-2 virus, has prompted global health concerns. In response, researchers have been conducting investigations on active compounds in plants that may hold the potential to inhibit the proliferation of the virus. The aim of this study was to simulate and predict structural interactions of selected compounds isolated from 28 endemic plants of Bosnia and Herzegovina against the main protease (Mpro), papain-like protease (PLpro), RNA-dependent RNA polymerase (RdRp), spike glycoprotein and uridylate-specific endoribonuclease (NendoU) of SARS-CoV-2. The majority of compounds, especially hesperidin, showed great binding affinity to the target proteins. The highest affinity for Mpro was observed for genistein and hesperidin, while in terms of structural interactions, both compounds achieved interactions of interest. Hesperidin and luteolin were the compounds with the highest binding affinity for PLpro, but no significant interactions were observed. For RdRp, hesperidin and quercetin showed the highest binding affinity, where both compounds formed interactions of interest. Hesperidin and fisetin were the compounds with the highest binding affinity for spike glycoprotein, and both compounds achieved significant interactions. The highest affinity for NendoU was obtained for hesperidin and isorhamnetin, where both compounds formed interactions of interest. Although these findings appear encouraging, further research is needed, which includes in vitro and in vivo assessments, along with clinical trials, to provide evidence for the potential therapeutic uses of these plants.

Molecular modeling and biological investigation of novel s-triazine linked benzothiazole and coumarin hybrids as antimicrobial and antimycobacterial agents

A novel series of s-triazine linked benzothiazole and coumarin hybrids (6a-6d, 7a-7d, and 8a-8d) were synthesized and characterized by IR, NMR, and mass spectrometry. The compound’s in vitro antibacterial and antimycobacterial activities were also evaluated. Remarkable antibacterial activity with MIC in the range of 12.5–62.5 μM and antifungal activity of 100–200 μM were demonstrated by in vitro antimicrobial analysis. Compounds 6b, 6d, 7b, 7d, and 8a strongly inhibited all bacterial strains, while 6b, 6c, and 7d had good to moderate efficacy against M. tuberculosis H37Rv. Synthesized hybrids are observed in the active pocket of the S. aureus dihydropteroate synthetase enzyme, according to a molecular docking investigations. Among the docked compounds, 6d had a strong interaction and a greater binding affinity, and the dynamic stability of protein-ligand complexes was examined using molecular dynamic simulation with various settings at 100 ns. The proposed compounds successfully maintained their molecular interaction and structural integrity inside the S. aureus dihydropteroate synthase, according to the MD simulation analysis. These in silico analyses supported the in vitro antibacterial results of compound 6d, which demonstrated outstanding in vitro antibacterial efficacy against all bacterial strains. In the quest for new antibacterial drug-like molecules, compounds 6d, 7b, and 8a have been identified as promising lead compounds. Communicated by Ramaswamy H. Sarma

Pharmacokinetics of Lixisenatide, a GLP-1 Receptor Agonist, Determined by a Novel Liquid Chromatography–Tandem Mass Spectrometry Analysis in Rats

Because of its greater binding affinity and longer half-life than native glucagon-like peptide-1 (GLP-1), the GLP-1 receptor agonist lixisenatide is commonly used to treat type 2 diabetes mellitus. This study aimed to establish a simple and robust liquid chromatography–tandem mass spectrometry (LC–MS/MS) approach for lixisenatide for in vivo pharmacokinetic investigation. Methanol-based protein precipitation with formic acid was exploited for plasma sample extraction, using esomeprazole as the internal standard. Gradient elution with 0.1% formic acid in distilled water and acetonitrile was utilized for chromatographic separation. Mass spectrometry was used to monitor the MRM transition at m/z 810.8 → 129.2 for lixisenatide. In rat plasma, lixisenatide had a lower limit of quantification of 10 ng/mL. The LC–MS/MS was applied to describe the pharmacokinetics of lixisenatide in rats following intravenous and subcutaneous dosing. The average half-life of lixisenatide was 0.37 ± 0.06 h after intravenous injection. The estimated subcutaneous bioavailability of lixisenatide was 2.17%. This LC–MS/MS analysis might be relevant in future research to create novel dosage formulations of lixisenatide and other GLP-1 receptor agonists with optimal therapeutic effectiveness.

Integrated molecular and quantum mechanical approach to identify novel potent natural bioactive compound against 2′-O-methyltransferase (nsp16) of SARS-CoV-2

With the advent of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) outbreak, efforts are still in progress to find out a functional cure for the infection. Among the various protein targets, nsp16 capping protein is one of the vital targets for drug development as it protects the virus against the host cell nucleases and evading innate immunity. The nsp16 protein forms a heterodimer with a co-factor nsp10 and triggers 2′-O-methyltransferase activity which catalyzes the conversion of S-adenosyl methionine into S-adenosyl homocysteine. The free methyl group is transferred to the 2′-O position on ribose sugar at the 5′ end of mRNA to form the cap-1 structure which is essential for replication of the virus and evading the innate immunity of the host. In this study, we identify a potential lead natural bioactive compound against nsp16 protein by systematic cheminformatic analysis of more than 144k natural compounds. Virtual screening, molecular docking interactions, ADMET profiling, molecular dynamics (MD) simulations, molecular mechanics-generalized born surface area (MM-GBSA), free energy analysis and density functional theory analysis were used to discover the potential lead compound. Our investigation revealed that ZINC8952607 (methyl-[(6-methyl-2,3,4,9-tetrahydro-1H-carbazol-1-yl)aminomethyl]BLAHone) has the greatest binding affinity and best pharmacokinetic parameters due to presence of carbazol and BLAHone (biaryl moiety). Further, time-dependent MD simulation analysis substantiates the stability and rigidness of nsp16 protein even after interaction with the lead compound. We believe that the compound ZINC8952607 might establish as a novel natural drug candidate against CoVID-19 infection. Communicated by Ramaswamy H. Sarma

Quantum Chemical Approaches to the Encapsulation of Parathion, Chlorpyrifos and Coumaphos by Armchair and Zigzag Boron Nitride Nanotubes Doped with Aluminum

Boron nitride nanotubes have been widely used as drug delivery vehicles and for the controlled release of targeted therapeutic drugs. In this study, we calculated the encapsulation efficiencies of three organophosphorus pesticides, parathion, chlorpyrifos, and coumaphous, using quantum chemical methods. The results show that the encapsulation energy of zigzag BNNT(20,0) is lower than that of armchair BNNT(12,12) to encapsulate parathion. Al doping helps to decrease the encapsulation energy and Al-doped zigzag BNNT(20,0) + parathion has the greatest binding affinity. In addition, the energy gap of armchair BNNT(12,12) encapsulating organophosphorus pesticides changed significantly. Al doping reduces the band gap of boron nitride nanotubes. Al-doped armchair BNNT(12,12) has the strongest electron-accepting ability and is a promising sensor material.

Abstract 6267: A novel and potent EZH1/2 dual inhibitor, HM97662 demonstrates antitumor activity in T-cell lymphoma

Abstract Chromatin remodeling is a crucial process for transcriptional regulation, of which dysregulation is often observed in various human cancers. The enhancer of zeste homology 2 (EZH2) and its homolog EZH1 are catalytic subunits of polycomb repressive complex 2 (PRC2), which trimethylate histone H3 at lysine 27 (H3K27me3) to repress transcription of its target genes. Although methlytransferase activity of PRC2 is mainly contributed by EZH2, EZH1 also conducts a compensatory role to maintain tri-methylation of H3K27. EZH1 also directly binds to chromatin and modulates its condensation. Recent studies have suggested that EZH1 as well as EZH2 played a critical role in T-cell lymphoma such as ATL/L and PTCL, which had high innate EZH1 and increased EZH2 expression upon acquisition of their malignancy. Consequently, dual inhibition of EZH1/2 might induce higher expression of downstream tumor suppressor genes than blocking EZH2 alone, expecting greater activity as an anti-cancer therapy. Herein, we presented a novel and potent EZH1/2 dual inhibitor, HM97662, which simultaneously inhibited the methyltransferase activity of both EZH1 and EZH2 with 2.1 and 16 nM of IC50, respectively. Surface plasmon resonance (SPR) assay verified that HM97662 had great binding affinity on EZH1-EED-SUZ12 complex as well as EZH2-EED-SUZ12 complex than competitors. HM97662 also showed broad and strong antiproliferative activity against various T-cell lymphoma cell lines. Representatively in HH cells, HM97662 not only suppressed global tri-methylation of H3K27, but also dose-dependently increased protein expression levels that modulate cell cycle or cancer cell apoptosis. HM97662 exhibited increment of CDKN1A (p21) proteins involved in cell cycle arrest and induced protein levels of cleaved caspase-3 as well as PARP. Additionally, resulting induction of apoptosis by HM97662 in HH cells was confirmed by TUNEL staining. Mechanistically, HM97662 increased mRNA expression of several target genes inducing cell cycle arrest and apoptosis in gene panel assay performed in HH cells. Two cell cycle repressor, CDKN2A (p16) and CDKN1C (p57), and a pro-apoptotic marker, BTG-2, were dose-dependently increased by HM97662. We further conducted chromatin accessibility assay and identified that the chromatin structures of them were loosened and highly transcribed after the treatment of HM97662. Based our in vitro pharmacology data, we evaluated an antitumor activity of HM97662 in EZH1/2 co-expressed HuT-102 T-cell lymphoma cell mouse xenograft model, and daily oral dosing of HM97662 showed potent tumor growth inhibition. In conclusion, the present studies demonstrated that HM97662 has promising prospective for the treatment of patients with T-cell lymphoma. Given that the necessity of new treatment options on T-cell lymphoma, it is urgent to assess the effectiveness of HM97662 in further clinical trials. Citation Format: Jooyun Byun, Seung Hyun Jung, Yu-Yon Kim, Miyoung Lee, Gunwoo Lee, Heesun Moon, Eun Young Lee, Junghwa Park, Seon Yeong Han, Young Gil Ahn, Young Hoon Kim, Kwee Hyun Suh. A novel and potent EZH1/2 dual inhibitor, HM97662 demonstrates antitumor activity in T-cell lymphoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6267.

Abstract 408: Novel growth hormone receptor antagonist with discrete PEGylations markedly improve response to therapy in human pancreatic and bile duct cancers in mice

Abstract Here we present a new and improved GHR antagonist candidate (compound G) with novel discrete PEGylations exhibiting marked efficacy in synergizing anti-cancer therapies. The American Cancer Society estimates the 5-year survival rates of pancreatic adenocarcinoma (PAAD) and bile duct cancer/cholangiocarcinoma (CHOL) to be a dismal 10% in 2022, indicating an urgent need for new therapeutic approaches for both of these devastating diseases. Several human cancers, including PAAD and CHOL express two validated oncological targets - the growth hormone receptor (GHR) and the insulin-like growth factor 1 receptor (IGF1R), as well as their cognate ligands growth hormone (GH) and insulin-like growth factor 1 (IGF1), respectively. Our laboratory had pioneered the discovery of pharmacologic antagonists for the growth hormone receptor (GHR), including the only approved and prescribed GHR antagonist, pegvisomant (Pfizer), prescribed for acromegaly (a condition of excess GH due to a hypersecreting pituitary adenoma). GHR antagonists not only inhibit the effects of GH but also robustly suppress the production of IGF1. The novel GHR antagonist compound G, that we present here, has a greater binding affinity than Pegvisomant to the GHR. And unlike Pegvisomant, compound G binds to the prolactin receptor as well as the GHR. A large body of research by us and others have earlier shown that the covert actions of tumoral GHR and IGF1R activation collectively drive mechanisms of therapy resistance including, but not limited to, active multidrug efflux, activation of epithelial-to-mesenchymal transition (EMT), resistance to apoptosis, increased fibrosis, increase of cancer stem cells, tumor angiogenesis and senescence. Our novel discrete PEGylated GHR antagonist, compound G, alone exhibits significant suppression of cancer cell viability, invasive potential, and drug efflux rate in PAAD and CHOL cell cultures. In Nude mice with human PAAD or CHOL xenografts, GHR inhibition by compound-G markedly sensitized the tumors towards both low and high doses of gemcitabine (for PAAD) or gemcitabine-cisplatin (for CHOL) treatments. Compound-G in combination with chemotherapy resulted in markedly reduced tumor sizes in all CHOL and PAAD mice and almost complete tumor clearance in 40% of the mice with PAAD. In the GHR antagonist treated tumors, significantly lower expression of ABC transporters (Abcb1, Abcg5, Abcc2, Abcg2), EMT mediators (Cdh2, Zeb1, Snai2, Epas1) and apoptosis marker (Bcl2) genes compared to control were observed - accounting for the molecular mechanisms underlying the observed effects. Collectively, our data provide a robust pre-clinical validation for treatment regimens combining GHR antagonist and chemotherapy leading to highly efficacious tumor clearance. Citation Format: Reetobrata Basu, John J. Kopchick. Novel growth hormone receptor antagonist with discrete PEGylations markedly improve response to therapy in human pancreatic and bile duct cancers in mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 408.

Abstract 5343: Evaluating the screened polyphenolics molecules as potential chemosensitizers against the oncogenic signalling proteins in lung cancer

Abstract Background: Lung cancer is the most commonly diagnosed cancer globally. There are numerous agonists and antagonistic treatment interventions available for therapy. However, all of them possess different detrimental consequences in addition to resistance and remission in sufferers. Medicinal plants have traditionally been employed to treat a wide range of ailments and problems. Methodology: A total of 550 flavonoids and small molecules retrieved from literature and databases (Chembl, and PubChem) were analysed for molecular docking, biological activities, ADME, and drug-like properties through SwissADME and PreADMET, Molecular Simulation, and cytotoxicity studies. Simultaneously, review literature is employed to identify the novel protein targets viz. ROS1 (3ZBF) & EGFR (6DUK). Additionally, the crystal structures of the proteins were obtained from PDB and their domains and active sites were predicted using ScanProsite, MOTIF Search, metaPOCKET2, and PrankWeb tools. Finally, the proteins and ligands were energy-minimized using Chimera and open babel tool and subjected to virtual screening and molecular docking using GOLD. Furthermore, the molecular simulation was performed at 100ns by utilizing GROMACS, and cytotoxicity studies were performed by employing the A549 Lung cancer cell line. Results: Diosmin (64.57) and Hesperidin (60.98) had the greatest binding affinity with the identified targets, i.e., ROS1 (3ZBF) and EGFR(6DUK) respectively against Lung Cancer, and remained stable during 100ns of molecular simulation. Furthermore, we have performed cytotoxicity studies on A549 cell lines that show both the compounds have significant anticancer activity and do not affect healthy cells in the range of, i.e., Diosmin (500 µg/ml to 0.024 µg/ml.) and Hesperidin (500 µg/ml to 125 µg/ml). Conclusion: After the analyses, we can conclude that it has the potential to be exploited as a novel therapeutic agent in the treatment of lung cancer.Keywords: Diosmin, Hesperidin, In-silico, A549, Lung Cancer, Simulation Citation Format: Swati Arora, Sumit Sheoran, Tanmayee Basu, Naidu Subbarao, Atul Kumar Upadhyay, Sugunakar Vuree. Evaluating the screened polyphenolics molecules as potential chemosensitizers against the oncogenic signalling proteins in lung cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5343.

A combined DFT and molecular docking study on novel tricarbonylrhenium(I) complexes bearing mono- and bivalent benzenesulfonamide scaffolds as human carbonic anhydrase IX and XII inhibitors

The aim of this study was to determine the reactivity and stability properties of a novel series of tricarbonylrhenium(I) complexes with 1,2,3-triazole-pyridine based on monovalent and bivalent benzenesulfonamide ligands as well as analyze their binding affinity within the active site of carbonic anhydrases IX and XII, using DFT and molecular docking. In the first setup, the DFT method was used to investigate the molecular geometries and vibrational frequencies of ligands and their Re(I) complexes. The basis set used was B3LYP/LanL2DZ-ECP/6-31G(d). The optimized geometric parameters (bond lengths and bond angles) and vibrational frequencies agree well with the experimental results. TD-DFT calculations were also performed to determine the influence of the methylene linker (-CH2-) on the bands found in the electronic spectra of these compounds. Global reactivity indices calculated from the energies of frontier molecular orbitals were used to assess their reactivity and stability properties (FMOs). Molecular electrostatic potential maps of the molecules were calculated to provide information about their chemical reactivity and to explain their intermolecular interactions. The computational data showed that bivalent ligands and their bivalent Re(I) complexes are more reactive than their monovalent counterparts. The second setup involves docking studies of the compounds within the active sites of the receptors (6QN2 for hCA IX and 1JD0 for hCA XII), which are performed using the Molegro Virtual Docker and AutoDock 4.2 programs, respectively. Our results indicate that bivalent ligands and their associated Re(I) complexes have a greater binding affinity than monovalent ligands and their associated Re(I) complexes. Compounds bis-L1 and bis-L2 exhibited the best affinity potential for hCA IX and hCA XII inhibition, making them the most promising candidates among those examined.

Remdesivir-induced conduction abnormalities: A molecular model-based explanation.

Purpose: Remdesivir use in COVID-19 is associated with cardiac conduction abnormalities from unclear mechanisms. A proposed mechanism is the bioaccumulation of the intermediate metabolite GS-441524 resulting in exogenous activation of cardiac adenosine A1 due to the structural similarity between adenosine and GS-441524. The prolonged half-life of GS-441524 can result in sustained activation of adenosine A1 receptors. In this study, we used molecular modeling of adenosine, GS-441524 and the adenosine A1 receptor to assess the potential mechanistic association of the proposed mechanism. Methods: Adenosine and GS-441524 structures were acquired from the PubChem database. Ligand docking was carried out using UCSF Chimera. Models were chosen based on greatest binding affinity and minimum root mean square deviation. Figures of resulting structural models were prepared using UCSF Chimera or PyMOL 2.3.5. Results: By modeling the interaction between the A1 G protein complex and both adenosine and GS-441524, we found that the proposed mechanism of exogenous A1 receptor activation is feasible based on docking compatibility. Conclusion: The proposed mechanism of exogenous cardiac A1 receptor activation from bioaccumulation of GS-441524 as a cause of observed cardiac conduction abnormalities with the use of remdesivir in COVID-19 is viable. Further studies are needed to assess causality.

Mechanistic insights of Cucumis melo L. seeds for gastrointestinal muscle spasms through calcium signaling pathway–related gene regulation networks in WGCNA and in vitro, in vivo studies

BackgroundIn addition to the nutritional benefits of Cucumis melo L., herbalists in Pakistan and India employ seeds to treat various ailments. This study aimed to determine the regulatory role of C. melo seeds in calcium-mediated smooth muscle contraction. MethodsWe identified and quantified the phytochemicals of C. melo with LC ESI–MS/MS and HPLC, then conducted in vitro and in vivo tests to confirm the involvement in smooth muscle relaxation. Then, diarrhea-predominant irritable bowel syndrome gene datasets from NCBI GEO were acquired, DEGs and WGCNA followed by functional enrichment analysis. Next, molecular docking of key genes was performed. ResultsThe quantification of C. melo seeds revealed concentrations of rutin, kaempferol, and quercetin were 702.38 μg/g, 686.29 μg/g, and 658.41 μg/g, respectively. In vitro experiments revealed that C. melo seeds had a dose-dependent relaxant effect for potassium chloride (80 mM)–induced spastic contraction and exhibited calcium antagonistic response in calcium dose-response curves. In in vivo studies, Cm.EtOH exhibited antidiarrheal, antiperistaltic, and antisecretory effects. The functional enrichment of WGCNA and DEGs IBS-associated pathogenic genes, including those involved in calcium-mediated signaling, MAPK cascade, and inflammatory responses. MAPK1 and PIK3CG were identified as key genes with greater binding affinity with rutin, quercitrin, and kaempferol in molecular docking. ConclusionsThe bronchodilator and antidiarrheal effects of C. melo were produced by altering the regulatory genes of calcium-mediated smooth contraction.