HSPiP, Computational, and Thermodynamic Model-Based Optimized Solvents for Subcutaneous Delivery of Tolterodine Tartrate and GastroPlus-Based In Vivo Prediction in Humans: Part I.

Tolterodine tartrate (TOTA) is associated with adverse effect, high hepatic access, varied bioavailability, slight aqueous solubility, and short half-life after oral delivery. Hansen solubility parameters (HSP, HSPiP program), experimental solubility (T = 298.2 to 318.2 K and p = 0.1 MPa), computational (van't Hoff and Apelblat models), and thermodynamic models were used to the select solvent(s). HSPiP predicted PEG400 as the most suitable co-solvent based on HSP values (δd = 17.88, δp = 4.0, and δh = 8.8 of PEG400) and comparable to the drug (δd = 17.6, δp = 2.4, and δh = 4.6 of TOTA). The experimental mole fraction solubility of TOTA was maximum (xe = 0.0852) in PEG400 confirming the best fit of the prediction. The observed highest solubility was attributed to the δp and δh interacting forces. The activity coefficient (ϒi) was found to be increased with temperature. The higher values of r2 (linear regression coefficient) and low RMSD (root mean square deviation) indicated a good correlation between the generated "xe" data for crystalline TOTA and the explored models (modified Apelblat and van't Hoff models). TOTA solubility in "PEG400 + water mixture" was endothermic and entropy-driven. IR (immediate release product) formulation can be tailored using 60% PEG400 in buffer solution for 2 mg of TOTA in 0.25 mL (dosing volume). The isotonic binary solution was associated with a pH of 7.2 suitable for sub-Q delivery. The approach would be a promising alternative with ease of delivery to children and aged patients.

COVID-19 vaccine hesitancy among adults in India: A primary study based on health behavior theories and 5C psychological antecedents model.

Despite the significant success of India's COVID-19 vaccination program, a sizeable proportion of the adult population remains unvaccinated or has received a single dose of the vaccine. Despite the recommendations of the Government of India for the two doses of the COVID-19 vaccine and the precautionary booster dose, many people were still hesitant towards the COVID-19 full vaccination. Hence, this study aimed to identify the primary behavioral and psychological factors contributing to vaccine hesitancy. Cross-sectional data was collected via a multi-stage sampling design by using a scheduled sample survey in the Gorakhpur district of Uttar Pradesh, India, between 15 July 2022 to 30 September 2022. This study has utilized three health behavior models-the Health Belief Model (HBM), the Theory of Planned Behavior (TPB), and the 5C Psychological Antecedents of vaccination, and employed bivariate and multivariable binary logistic regression model to assess the level of vaccine hesitancy and predictive health behavior of the respondents. Results indicate that among the constructs of the HBM and 5C Antecedents models, "perceived benefits", "confidence" and "collective responsibility" showed a lesser likelihood of COVID-19 vaccine hesitancy. However, in the TPB model constructs, a 'negative attitude towards the vaccine' showed a four times higher likelihood of COVID-19 vaccine hesitancy. From the future policy perspective, this study suggested that addressing the issue of 'negative attitudes towards the vaccine' and increasing the trust or confidence for the vaccine through increasing awareness about the benefits of the vaccination in India may reduce vaccine hesitancy.

Potentially active aspirin derivative to release nitric oxide: In-vitro, in-vivo and in-silico approaches.

The series of newer salicylate derivatives incorporating nitroxy functionality were synthesized and evaluated for their potential effect in gastrointestinal (GI) related toxicity produced by aspirin. The synthesized compounds (5a-j) were subjected to %NO (nitric oxide) release study, in-vitro anti-inflammatory potential, % inhibition of carrageenan-induced paw edema and the obtained results were validated by in-silico studies including molecular docking, MD simulations and in-silico ADME (absorption, distribution, metabolism, and elimination) calculations. Compounds 5a (20.86 %) and 5g (18.20 %) displayed the highest percentage of NO release in all the tested compounds. Similarly, 5a and 5h were found to have (77.11 % and 79.53 %) &(78.56 % and 66.10 %) inhibition in carrageenan induced paw edema in animal mode which were relatively higher than ibuprofen (standard used). The obtained results were validated by molecular docking and MD simulations studies. The molecular docking study of 5a and 5h revealed that docking scores were also obtained in very close proximity of -8.35, -9.67 and -8.48 for ibuprofen, 5g and 5h respectively. In MD simulations studies, the calculated lower RMSD (root mean square deviation) values 2.8 Å and 5.6 Å for 5g and 5h, respectively indicated the stability of ligand-protein complexes. Similarly lower RSMF (root mean square fluctuation) values indicated the molecules remained in the active pocket throughout the entire MD simulations run. Further, in-silico ADME calculations were determined and all compounds obey the Lipinski's rule of five and it was predicted that these molecules would be orally active without any serious toxic effect.

Karanjin, A Promising Bioactive Compound Possessing Anti-cancer Activity against Experimental Model of Non-small Cell Lung Cancer Cells.

AIMS: The aim of this study is to isolate the Millettia pinnata (Karanj) leaf extract for pure compound with anticancer properties and to study the molecular target of the isolates in non-small cell lung cancer cell lines. BACKGROUND: In our earlier research Millettia pinnata leaf extract has demonstrated potential anticancer activities. Thus, in pursuit of the bioactive compounds, the most potential active extract from our previous study was purified. Furthermore, the anticancer properties of the isolated compound karanjin was studied and aimed for apoptosis and restraining growth. METHODS: A novel method was developed through column chromatography for isolation and purification of the compound karanjin from leaf chloroform extract. The purified component was then characterised using FTIR, mass spectrometry, and NMR. An MTT-based cytotoxicity assay was used to analyse cell cytotoxicity, whereas fluorescence staining was used for apoptosis and reactive oxygen species inhibition quantification. Furthermore, the real-time PCR assay was used to determine the molecular mechanism of action in cells causing cytotoxicity induced by karanjin dosing. RESULTS: The anticancer activity of karanjin in A549 cell line exhibited prominent activity revealing IC50 value of 4.85 µM. Conferring the predicted molecular pathway study, karanjin restrains the proliferation of cancer cells through apoptosis, which is controlled by extrinsic pathway proteins FAS/FADD/Caspases 8/3/9. Downregulation of KRAS and dependent gene expression also stopped cell proliferation. CONCLUSION: Karanjin has been identified as a compound with potential effect in non-small cell lung cancer cells. Molecular mechanism for apoptosis and inhibition of reactive oxygen species induced through H2O2 were observed, concluding karanjin have medicinal and antioxidant properties.

Status and correlates of non-communicable diseases among children and adolescents in slum and non-slum areas of India's four metropolitan cities.

The emergence of non-communicable diseases (NCDs) in childhood poses a serious risk to a healthy adult life. The present study aimed to estimate the prevalence of NCDs among children and adolescents in slums and non-slums areas of four metropolitan cities of India, and in rural areas of the respective states The study further, investigated the effect of the place residence as slum vs. non-slum and other risk factors of the NCDs. Nationally representative data from the Comprehensive National Nutrition Survey (CNNS) was used.. Estimates were based on children (5-9 years) and adolescents (10-19 years) for whom biomarkers predicting diabetes, high total cholesterol, high triglycerides and hypertension were determined. Weight, height and age data were used to calculate z-scores of the body mass index. Overweight and obesity was higher in urban areas than in rural areas among children and adolescents. Regional differences in the prevalence of diseases were observed; children in Delhi and Chennai had a higher likelihood of being diabetic while children in Kolkata were at a greater risk of high total cholesterol and high triglycerides. The risk of hypertension was strikingly high among non-slum children in Delhi. Children from slums were at a higher risk of diabetes compared to the children from non-slums, while children and adolecents from non-slums were at a greater risk of high triglycerides and hypertension respectively than their counterparts from slums. Male children and adolecents had a higher risk of diabetes and high cholesterol. Screening of children for early detection of NCDs should be integrated with the already existing child and adolescent development schemes in schools and the community can help in prevention and control of NCDs in childhood.

Exploration of spatial clustering in maternal health continuum of care across districts of India: A geospatial analysis of demographic and health survey data.

INTRODUCTION: The continuum of care (CoC) throughout pregnancy, delivery and post-delivery has recently been highlighted as an integrated intervention programme for maternal, new-born, and child health. Existing literature suggests the importance of continuum of care (CoC) for improved maternal and child health outcomes. However due to unavailability of data at the lowest administrative levels, literature on spatial pattern of uptake of full CoC is lacking. The present study attempts to focus on the spatial analysis of CoC in maternal health care in India. DATA AND METHODS: The study is based on the fourth round of National Family Health Survey data conducted in 2015-16 in India. The outcome variable used is maternal health continuum of care which includes- at least 4 ANC visits, delivery through skilled birth attendant and postnatal check-up within 48 hours of delivery. Univariate and bivariate Local Indicator of Spatial Association (LISA) maps have been generated to show the spatial pattern of CoC across 640 districts in India. We also employed spatial regression techniques to explore the determinants of CoC. FINDINGS: Percentage of women who followed full CoC was observed to be least for East Kameng (0.0%) district of Arunachal Pradesh and highest in North Goa district (90.4%). Majority of districts where uptake of full CoC was more than 80 percent were found concentrated in southern region on India. Equivalently, findings indicated a strong spatial clustering of full CoC with high-high clusters mostly concentrated in southern districts. Low-low district clusters are concentrated in the states of Uttar Pradesh, Bihar and Madhya Pradesh. For complete CoC the global Moran's I is 0.73 indicating the spatial dependence. The spatial regression analysis suggested that modern contraceptive use, meeting with health worker, urbanization and secondary or above education for women have positive impact on the utilisation of CoC. CONCLUSION: The spatial pattern indicates district level clustering in uptake of CoC among women. The study suggests policymakers and stakeholders to implement comprehensive interventions at sub-regional levels for ensuring the completion of CoC for women which acts as a preventive measure for adverse outcomes such as-maternal and child mortality.

Status and correlates of micronutrient deficiencies in slum and non-slum areas of India's four metropolitan cities: Investigation from CNNS.

Micronutrient malnutrition is an emerging public health concern globally. It affects people of all ages and socioeconomic groups; however, the most marginalized are the worst affected. Using data from the Comprehensive National Nutrition Survey 2016-18, we determined the magnitude of deficiencies (of iron, zinc, vitamin A, folate, vitamins B12 and D) among children and adolescents (1-19 years of age) living in four metropolitan cities of India. Separate estimates by residence in slum and non-slum areas were derived for pre-school and school-aged children and adolescents. The association between each micronutrient deficiency (MND) and place of residence, exposure to progarmmes, socioeconomic, and demographic variables was assessed using Poisson regression. Of all children in the sample, at least seven out of 10 children suffered from some kind of MND. Anaemia was prevalent among all children but at different levels among various age-groups. Folate deficiency was highly prevalent among children in slums whereas deficiencies of vitamin D and zinc were more prevalent among non-slum children. Dietary diversity reduced the risk of deficiencies- Vitamin A in children 1-9, anaemea in 1-4 age, folate in children 5-19. Exposure to government-sponsored nutrition programmes such as mid-day meal, and IFA did not show any significant effect on reducing deficiencies. However, adolescents exposed to IFA supplementation programmes were less likely to be folate deficient. Overall, government schemes that have been running for decades, and intensified lately, are yet to show noticeable positive effect on micronutrient status of children. Nevertheless, differential estimates by slum/non-slum residence and by age-groups calls for devising different strategies for different sub-groups to address the issue of MNDs among children and adolescents. Nutrition education not only for slum residents but also for those from non-slum areas is an urgent need to check the spread of MNDs.

Development and validation of a questionnaire to evaluate the knowledge, attitude and practices regarding travel medicine amongst physicians in an apex tertiary hospital in Northern India.

OBJECTIVES: Travel medicine focuses primarily on pre-travel preventive care and the conditions and diseases acquired during or after travel. There is a paucity of validated tools to assess the knowledge, attitude and practises of physicians with regard to travel medicine. We attempted to develop a tool to assess existing expertise among Medicine and Infectious Diseases resident doctors with respect to travel medicine. METHODS: Item level content validity index (I-CVI) and scale level content validity index (S-CVI/Ave) were estimated for each of the items to establish the content validity. Refined measures of inter-rater agreement (Brennan and Prediger Agreement Coefficient and Gwet's Agreement Coefficient) were estimated for the tool. RESULTS: The final version of the questionnaire had satisfactory content validity (I-CVI > 0∙6 and S-CVI/Ave > 0∙9) and possessed high agreement among the raters (Brennan and Prediger AC > 0∙7, p < 0∙01 and Gwet's AC > 0∙8, p < 0∙01) with regard to necessity, clarity and relevance of the scale. CONCLUSIONS: This tool covers a wide range of questions and is scientifically validated. The final version of the tool can be used largely for the assessment of knowledge, attitude and practices among medical practitioners. This is instrumental to build targeted intervention programs to enhance the knowledge regarding travel medicine among health care providers.

Selective adsorption of anionic dye from wastewater using polyethyleneimine based macroporous sponge: Batch and continuous studies.

Dyes are well known for their hazardous impacts on public health and the environment. Dye removal using monolithic adsorbents is an attractive approach for industrial applications and process design owing to their utilization in both static and dynamic adsorption experiments. In the present work, polyethyleneimine (PEI) based macroporous monolithic sponge (S100) was engineered by ice-templating method and used as an adsorbent. Both batch and continuous operations for dye removal were studied. The effect of various parameters such as pH, adsorbent amount, flow rate, influent dye concentration, and adsorbent bed height on adsorption performance of S100 was studied and modelled using Langmuir/Freundlich isotherms for static operations and Adam-Bohart/Thomas model in packed-bed column experiments. Under optimum conditions, the adsorbent showed a remarkably higher adsorption capacity towards CR (1666.67 mg/g), which is considerably higher than most PEI-based adsorbents. Amine groups in S100 offered exceptional selectivity for anionic Congo red (CR) against cationic Methylene blue (MB) dye (separation factor of 208 and 87 in absence and presence of sodium chloride, respectively). It can be easily regenerated in alkaline medium without a significant loss in percent adsorption capacity and shows good thermal and mechanical stability. Notably, in column studies, a relatively smaller percentage of unused bed height (32.3%) was observed with higher dye uptake for 16 mg S100 at flow rate 10 mL/h and inlet concentration 300 mg/L. Thus, the adsorbent displays an outstanding physiochemical characteristic, excellent selectivity for anionic dye, ease of regeneration and high adsorption performance in both batch and continuous studies.

Enhanced modelling and experimental validation of ultra-low pressure reverse osmosis membrane system for treatment of synthetic brackish water.

Water purification from brackish water sources has been acknowledged as one of the most promising ways to produce drinkable water in water-scarce areas. In this study, an ultra-low pressure reverse osmosis (ULPRO) membrane was numerically and experimentally investigated to produce drinking water by the removal of sodium chloride salt which provides further validation of the model from a practical perspective. An enhanced predictive model based on the Donnan-Steric Pore Model with dielectric exclusion (DSPM-DE) incorporating the osmotic effects was formulated in process simulation. The feed pressure and concentration were optimized as input variables and interaction between them was observed, while salt rejection and water recovery rate were taken as response attributes. The results obtained on the ULPRO membrane showed that the performance depends on the charge, steric, and dielectric effects. Furthermore, the enhanced model was validated with the experimental data attained from a laboratory-scale filtration system with good accuracy in the salt rejection and water recovery results. Comparing the enhanced DSPM-DE with the existing solution diffusion model reveals that the enhanced model predicts the membrane performance better and thereby qualifies itself as a reliable model for desalination of brackish water using ULPRO membrane.

Preparation and Evaluation of Silymarin-Loaded Solid Eutectic for Enhanced Anti-Inflammatory, Hepatoprotective Effect: In Vitro-In Vivo Prospect.

Solubility of phytochemicals is a major concern for drug delivery, permeability, and their biological response. However, advancements in the novel formulation technologies have been helping to overcome these challenges. The applications of these newer technologies are easy for commercialization and high therapeutic outcomes compared to conventional formulations. Considering these facts, the present study is aimed to prepare a silymarin-loaded eutectic mixture with three different ratios of Polyvinylpyrrolidone K30 (PVP K30) and evaluating their anti-inflammatory, and hepatoprotective effects. The preliminary phytochemical and characterization of silymarin, physical mixture, and solid dispersions suggested and successfully confirmed the formation of solid dispersion of silymarin with PVP K30. It was found that the solubility of silymarin was increased by 5-fold compared to pure silymarin. Moreover, the in vitro dissolution displayed that 83% of silymarin released within 2 h with 2.8-fold increase in dissolution rate compared to pure silymarin. Also, the in vivo study suggested that the formulation significantly reduced the carbon tetrachloride- (0.8620 ± 0.05034∗∗ for 1 : 3 ratio), paracetamol- (0.7300 ± 0.01517∗∗ for 1 : 3 ratio), and ethanol- (0.8100 ± 0.04037∗∗ for 1 : 3 ratio) induced hepatotoxicity in rats. Silymarin solid dispersion was prepared using homogenization methods that have prominent anti-inflammatory effect (0.6520 ± 0.008602∗∗ with 8.33%) in carrageenan-induced rat paw model.

Thermodynamic, Computational Solubility Parameters in Organic Solvents and In Silico GastroPlus Based Prediction of Ketoconazole.

The study aimed to select a suitable solvent capable to solubilize ketoconazole (KETO) and serve as a permeation enhancer across the skin. Experimental solubility and Hansen solubility parameters were obtained in ethanol, dimethyl sulfoxide (DMSO), ethylene glycol, oleic acid, span 80, limonene, eugenol, transcutol (THP), labrasol, and propylene glycol. Thermodynamic functional parameters and computational models (van't Hoff and Apelblat) validated the determined solubility in various solvents at T = 298.2 K to 318.2 K and P = 0.1 MPa. The HSPiP software estimated the solubility parameters in the solvents. The maximum mole fractional solubility values of KETO were found to be in an order as oleic acid (8.5 × 10-3) > limonene (7.3 × 10-3) > span 80 (6.9 × 10-2) > THP (4.9 × 10-2) > eugenol (4.5 × 10-3) at T = 318.2 K. The results of the apparent thermodynamic analysis confirmed that the dissolution rate was endothermic and entropy driven. The GastroPlus program predicted significantly high permeation of KETO (79.1%) in human skin from the KETO-THP construct as compared to drug solution (38%) and excellent immediate release from THP-solubilized construct (90% < 1 h). Hence, THP could be a better option for topical, transdermal, and oral formulation.

Continuum of maternal health care services and its impact on child immunization in India: an application of the propensity score matching approach.

Continuum of care throughout pregnancy, delivery and post-delivery has proved to be a critical health intervention for improving the health of mothers and their newborn children. Using data from the fourth wave of the National Family Health Survey (NFHS-4) conducted in 2015-16, this study examined the correlates of utilization of maternal health care services and child immunization following the continuum of care approach in India. The study also assessed whether the continuity in utilizing maternal health care services affects the immunization of children. A total of 33,422 survey women aged 15-49 were included in the analysis of maternal health care indicators, and 8246 children aged 12-23 months for the analysis of child immunization. The results indicated that about 19% of the women had completed the maternal health continuum, i.e. received full antenatal care, had an institutional delivery and received postnatal care. Women with a higher level of education and of higher economic status were more likely to have complete continuum of care. Continuity of maternal health care was found to be associated with an increase in the immunization level of children. It was observed that 76% of the children whose mothers had complete continuum of care were fully immunized. Furthermore, the results from propensity score matching revealed that if mothers received continuum of care, the chance of their child being fully immunized increased by 17 percentage points. The results suggest that promotion of the continuum of maternal health care approach could help reduce not only the burden of maternal deaths in India, but also that of child deaths by increasing the immunization level of children.

Quantum chemical insight into molecular structure, NBO analysis of the hydrogen-bonded interactions, spectroscopic (FT-IR, FT-Raman), drug likeness and molecular docking of the novel anti COVID-19 molecule 2-[(4,6-diaminopyrimidin-2-yl)sulfanyl]-N-(4-fluorophenyl)acetamide - dimer.

Novel antiviral active molecule 2- [(4,6-diaminopyrimidin-2-yl)sulfanyl]-N-(4-fluoro- phenyl)acetamide has been synthesised and characterized by FT-IR and FT-Raman spectra. The equilibrium geometry, natural bond orbital calculations and vibrational assignments have been carried out using density functional B3LYP method with the 6-311G++(d,p) basis set. The complete vibrational assignments for all the vibrational modes have been supported by normal coordinate analysis, force constants and potential energy distributions. A detailed analysis of the intermolecular interactions has been performed based on the Hirshfeld surfaces. Drug likeness has been carried out based on Lipinski's rule and the absorption, distribution, metabolism, excretion and toxicity of the title molecule has been calculated. Antiviral potency of 2- [(4,6-diaminopyrimidin-2-yl)sulfanyl]-N-(4-fluoro-phenyl) acetamide has been investigated by docking against SARS-CoV-2 protein. The optimized geometry shows near-planarity between the phenyl ring and the pyrimidine ring. Differences in the geometries due to the substitution of the most electronegative fluorine atom and intermolecular contacts due to amino pyrimidine were analyzed. NBO analysis reveals the formation of two strong stable hydrogen bonded N-H···N intermolecular interactions and weak intramolecular interactions C-H···O and N-H···O. The Hirshfeld surfaces and consequently the 2D-fingerprint confirm the nature of intermolecular interactions and their quantitative contributions towards the crystal packing. The red shift in N-H stretching frequency exposed from IR substantiate the formation of N-H···N intermolecular hydrogen bond. Drug likeness and absorption, distribution, metabolism, excretion and toxicity properties analysis gives an idea about the pharmacokinetic properties of the title molecule. The binding energy -8.7 kcal/mol of the nonbonding interaction present a clear view that 2- [(4,6-diaminopyrimidin-2-yl)sulfanyl]-N-(4-fluoro- phenyl) acetamide can irreversibly interact with SARS-CoV-2 protease.

A Series of Ferulic Acid Amides Reveals Unexpected Peroxiredoxin 1 Inhibitory Activity with in†vivo Antidiabetic and Hypolipidemic Effects.

Insulin resistance is a major pathophysiological feature in the development of type 2 diabetes (T2DM). Ferulic acid is known for attenuating the insulin resistance and reducing the blood glucose in T2DM rats. In this work, we designed and synthesized a library of new ferulic acid amides (FAA), which could be considered as ring opening derivatives of the antidiabetic PPARγ agonists Thiazolidinediones (TZDs). However, since these compounds displayed weak PPAR transactivation capacity, we employed a proteomics approach to unravel their molecular target(s) and identified the peroxiredoxin 1 (PRDX1) as a direct binding target of FAAs. Interestingly, PRDX1, a protein with antioxidant and chaperone activity, has been implied in the development of T2DM by inducing hepatic insulin resistance. SPR, mass spectrometry-based studies, docking experiments and in vitro inhibition assay confirmed that compounds VIe and VIf bound PRDX1 and induced a dose-dependent inhibition. Furthermore, VIe and VIf significantly improved hyperglycemia and hyperlipidemia in streptozotocin-nicotinamide (STZ-NA)-induced diabetic rats as confirmed by histopathological examinations. These results provide guidance for developing the current FAAs as new potential antidiabetic agents.

The effects of aspirin gel and mouthwash on levels of salivary biomarkers PGE2, TNF-α and nitric oxide in patients with periodontal diseases.

Inflammation and its mediators have an important role in gingivitis and periodontitis. Prostaglandin is one of the eicosanoid involved in many chronic inflammatory diseases, including periodontal diseases. Aspirin irreversibly acetylates cyclooxygenase and inactivate this enzyme responsible for the production of PGE2 that mediates pain and inflammation. The aim of the study was to prepare aspirin gel and mouthwash in 1% concentration and use it in patients with periodontal diseases during the non-surgical periodontal treatment and to assess its anti-inflammatory effects on salivary biomarkers PGE2, TNF-α, and nitric oxide. Thirty patients were divided into three treatment groups, standard treatment group, second received scaling and root planning with gel application of 1% aspirin, third received scaling and root planning followed by rinsing with 1% aspirin mouthwash. Results indicated that the levels of PGE2, TNF-α and nitric oxide in the groups of patients received gel treatment and mouthwash treatment was decreased to significant levels (p<0.001) as compared to the group of standard treatment. Aspirin gel was found to be more effective in reducing inflammatory biomarkers in contrast to aspirin mouthwash (p<0.001). We concluded from our study, that low concentration of aspirin oral preparations are highly active in reducing the inflammatory biomarkers associated with periodontal diseases.

CYP enzymes, expressed within live human suspension cells, are superior to widely-used microsomal enzymes in identifying potent CYP1A1/CYP1B1 inhibitors: Identification of quinazolinones as CYP1A1/CYP1B1 inhibitors that efficiently reverse B[a]P toxicity and cisplatin resistance.

Microsomal cytochrome P450 (CYP) enzymes, isolated from recombinant bacterial/insect/yeast cells, are extensively used for drug metabolism studies. However, they may not always portray how a developmental drug would behave in human cells with intact intracellular transport mechanisms. This study emphasizes the usefulness of human HEK293 kidney cells, grown in 'suspension' for expression of CYPs, in finding potent CYP1A1/CYP1B1 inhibitors, as possible anticancer agents. With live cell-based assays, quinazolinones 9i/9b were found to be selective CYP1A1/CYP1B1 inhibitors with IC50 values of 30/21 nM, and > 150-fold selectivity over CYP2/3 enzymes, whereas they were far less active using commercially-available CYP1A1/CYP1B1 microsomal enzymes (IC50, >10/1.3-1.7 µM). Compound 9i prevented CYP1A1-mediated benzo[a]pyrene-toxicity in normal fibroblasts whereas 9b completely reversed cisplatin resistance in PC-3/prostate, COR-L23/lung, MIAPaCa-2/pancreatic and LS174T/colon cancer cells, underlining the human-cell-assays' potential. Our results indicate that the most potent CYP1A1/CYP1B1 inhibitors would not have been identified if one had relied merely on microsomal enzymes.

Bergapten inhibits liver carcinogenesis by modulating LXR/PI3K/Akt and IDOL/LDLR pathways.

Oxysterol receptors LXRs (α and ß) are recently reported to be one of the novel and potential therapeutic targets in reducing cell proliferation and tumor growth in different system model. Activation of LXRs is correlated with modification of PI3K/Akt pathway. LXRs are also found to play a critical role in maintaining lipid homeostatais by regulating ABCA1, IDOL, SREBP1, LDLR and also certain lipogenic genes such as FASN and SCD1. In the present study a potential furanocoumarin, Bergapten (BeG) has been evaluated for its anticancer property on Hepatocellular Carcinoma (HCC) on LXR axis. The molecular docking analysis was carried out for BeG on LXR (α & ß) using Maestro tool and compared with reference ligands. This was followed by in vitro (HepG2 cell lines) and in vivo (on NDEA induced HCC in Wistar albino rats) anticancer evaluation of BeG. The docking results revealed polar and hydrophobic interactions of BeG with LXR (α,ß). The in vitro studies revealed the potential of BeG in lowering the accumulation of lipid droplets in HepG2 cells which was correlated with increase in LXR (α,ß) protein expressions. Furthermore, the in vivo studies demonstrated the potential of BeG in ameliorating the cancer induced alterations in body weight, liver weight and significant restoration of the changes in mRNA and protein expressions of LXR(α,ß), ABCA1, IDOL, SREBP1 and LDLR. BeG also modulated the expressions of PI3K, Akt and certain lipogenic genes like FASN and SCD1 and reduced the lipid droplets level in liver cancer cells. These results provide evidence and validates the critical role of BeG in maintaining the lipid homeostasis and justifies its anticancer potential against NDEA-induced HCC.

Taxifolin binds with LXR (α & ß) to attenuate DMBA-induced mammary carcinogenesis through mTOR/Maf-1/PTEN pathway.

AIM: 7,12-dimethylbenz(a)anthracene(DMBA), a PAH derivative initializes cascades of signaling events that alters a variety of enzymes responsible for lipid and glucose homeostasis resulting in enhanced availability and consumption of energy producing molecules for the development of carcinogenesis. 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoAR) is a key enzyme regulating the pathway of synthesis of cholesterol whereas liver-X-receptor (LXR) regulates lipid, carbohydrate metabolism in various malignancies including mammary carcinogenesis (MC). In this study Taxifolin (TAX), a potential flavanoid has been subjected to evaluate its anti-cancer potential on (MC). METHODS: We designed to screen the molecular docking analysis of TAX on LXRα, LXRß, HMG-CoAR, mTOR and PTEN using MAESTRO tool comparing with their reference ligands. MC was developed by the administration of DMBA in the air pouch (under the mammary fat pad) of the female Sprague-Dawley rats (55 days old). After 90 days of cancer induction, the chemotherapeutic potential of TAX was evaluated by administering TAX at different doses (10, 20 and 40 mg/kg b.w./day). Then western blot and RT-qPCR analysis were performed for determination of the protein and mRNA expressions respectively. RESULTS: The docking analysis revealed significant interaction with LXR (α&ß), HMG-CoAR, mTOR and PTEN. The docking results were validated with the enzyme inhibition assay using HMG-CoAR (EC 1.1.1.34). TAX inhibited the HMG-CoAR activity with an IC50 value of 97.54 ±â€¯2.5 nM whereas the reference molecule pavastatin revealed an IC50 value of 84.35 ±â€¯1.2 nM. Moreover, TAX modulated the energy regulation on DMBA-induced MC in SD-rats by significantly restoring the cancer-induced alterations in body weight, tumor growth and lipid, lipoproteins, lipid metabolizing enzymes and glycolytic enzymes. TAX interacted with LXRs, HMG-CoAR, metabolic enzymes and restored the altered metabolism that accelerates uncontrolled cell proliferation in MC. Moreover, TAX also altered the mRNA and protein expressions of HMG-CoAR, LXR (α,ß), Maf1, PTEN, phosphoinositide 3-kinase (PI3K), Akt, mTOR, fatty acid synthase (FASN) and acetyl-CoA carboxylase 1 (ACC1) in a dose dependent manner. CONCLUSION: These results validate the anti-cancer potential of TAX in DMBA-induced MC through LXR-mTOR/Maf1/PTEN axis.