Dual Anti-Inflammatory and Anticancer Activity of Novel 1,5-Diaryl Pyrazole Derivatives: Molecular Modeling, Synthesis, In Vitro Activity, and Dynamics Study.

A series of novel 1,5-diaryl pyrazole derivatives targeting the COX enzyme were designed by combined ligand and structure-based approach. The designed molecules were then further subjected to ADMET and molecular docking studies. Out of 34 designed compounds, the top-10 molecules from the computation studies were synthesized, characterized, and evaluated for COX-2 inhibition and anti-cancer activity. Initially, the target compounds were screened for the protein denaturation assay. The results of the top-five molecules T2, T3, T5, T6, and T9 were further subjected to in vitro COX-2 enzymatic assay and anti-cancer activity. As far as COX-2 inhibitory activity is considered, two compounds, T3 and T5, exhibited the half maximum inhibitory concentration (IC50) at 0.781 µM and 0.781 µM respectively. Further, the two compounds T3 and T5, when evaluated for COX-1 inhibition, exhibited excellent inhibitory activity with T3 IC50 of 4.655µM and T5 with IC50 of 5.596 µM. The compound T5 showed more significant human COX-2 inhibition, with a selectivity index of 7.16, when compared with T3, which had a selectivity index of 5.96. Further, in vitro anti-cancer activity was screened against two cancer cell lines in which compounds T2 and T3 were active against A549 cell lines and T6 was active against the HepG2 cell line. Stronger binding energy was found by comparing MM-PBSA simulations with molecular docking, which suggests that compounds T3 and T5 have a better possibility of being effective compounds, in which T5 showed higher binding affinity. The results suggest that these compounds have the potential to develop effective COX-2 inhibitors as anti-cancer agents.

Chemical Composition and Biological Activities of the Essential Oil from Citrus reticulata Blanco Peels Collected from Agrowastes.

Citrus fruits have a thick outer coat which is often discarded due to its low economic value and usually contributes to the waste. So this work focused on exploring the potential pharmacological properties of the discarded citrus peels. In the present study, we extracted the essential oil from peel wastes of Citrus reticulata Blanco (CREO) from the local market. The antioxidant, antibacterial, and anticancer properties of essential oil were evaluated. The CREO exhibited a strong antioxidant property with DPPH radical scavenging, ABTS radical scavenging, H2 O2 radical scavenging, Ferric reducing antioxidant power and for Lipid peroxidation inhibition respectively. Antibacterial properties of CREO was indicated against different pathogenic microbial strains like E. coli, P. aeruginosa, S. aureus, and S. enterica in terms of disc diffusion method and minimum inhibitory concentration (MIC). Further, anticancer properties studied on breast cancer cell lines MCF7 and MDA-MB-231 showed dose-dependent cytotoxicity with IC50 of 56.67±3.12 µg/mL and 76.44±2.53 µg/mL respectively. The GC-MS analysis of CREO revealed the presence of major compounds like S-limonene, α-pinene, α-myrcene, and cis-terpinene which might have played a significant role in strong antioxidant, antibacterial and anticancer properties. The study thus identified the potential health benefits of Citrus reticulata peel waste.

Antioxidant, Antimicrobial, Cytotoxicity, and Larvicidal Activities of Selected Synthetic Bis-Chalcones.

Plants are known to have numerous phytochemicals and other secondary metabolites with numerous pharmacological and biological properties. Among the various compounds, polyphenols, flavonoids, anthocyanins, alkaloids, and terpenoids are the predominant ones that have been explored for their biological potential. Among these, chalcones and bis-chalcones are less explored for their biological potential under in vitro experiments, cell culture models, and animal studies. In the present study, we evaluated six synthetic bis-chalcones that were different in terms of their aromatic cores, functional group substitution, and position of substitutions. The results indicated a strong antioxidant property in terms of DPPH and ABTS radical-scavenging potentials and ferric-reducing properties. In addition, compounds 1, 2, and 4 exhibited strong antibacterial activities against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella enteritidis. The disc diffusion assay values were indicative of the antibacterial properties of these compounds. Overall, the study indicated the antioxidant and antimicrobial properties of the compounds. Our preliminary studies point to the potential of this class of compounds for further in vivo investigation.

Withaferin A targets the membrane of Pseudomonas aeruginosa and mitigates the inflammation in zebrafish larvae; an in vitro and in vivo approach.

Infections due to multidrug-resistant Pseudomonas aeruginosa are prevalent among patients with cystic fibrosis. The emergence of antibiotic-resistant pathogens necessitated the development of novel low-risk natural antibacterial compounds. Herbal medicines are used from dates of the origin of mankind and still serve their purpose as therapeutic agents. We demonstrated the antibacterial activity of Withaferin A extracted from the traditional herb, ashwagandha or winter cherry (Withania somnifera). Withaferin A exhibits strong antibacterial activity against P. aeruginosa with a minimum inhibitory concentration of 60 µM and minimum bactericidal concentration of 80 µM. Results obtained from membrane stabilization assay and electron microscopic analysis showed that Withaferin A acts by damaging the cell membrane of P. aeruginosa. Additionally, we investigated oxidative stress and inflammatory response after Withaferin A treatment in P. aeruginosa infected zebrafish larvae model. The results indicate that the level of ROS, and its related lipid peroxidation and apoptosis were significantly reduced after treated with Withaferin A. Consequently, an increment in antioxidant enzymes level such as superoxide dismutase (SOD) and catalase (CAT) was observed. Macrophage localization experiment showed a smaller number of localized macrophages in zebrafish, which indicates the reduction in inflammatory response. In conclusion, Withaferin A could serve as an alternative natural product in the treatment of infections caused by P. aeruginosa.

Solanum pubescens Wild Fruits Essential Oil - A Golden Casket for its Antimicrobial and Anti-inflammatory Mediated Wound Healing Efficacy in Vertebrate Model Mus musculus.

BACKGROUND: Solanum pubescens Willd, growing wild in the hills of Rayadurg jurisdiction of Southwestern Andhra Pradesh, has gained significant attention among researchers for its diverse folkloric applications, existence of novel phytochemicals and leaf extracts which hold great medicinal promises. To date, the S. pubescens fruit's essential oil (SPO) has never been investigated. METHODS: The current research has been focused to evaluate the chemical composition of S. pubescens fruit essential oil through Gas Chromatography-Mass Spectrometry (GC-MS), followed by the investigation of antibacterial, antifungal, anti-inflammatory, analgesic and wound healing activities in appropriate models to uncover its biological potentials. Extraction of (Solanopuboil/SPO) from the fresh unripe fruits of Solanum pubescens was carried out in Buchner funnel and Whatman no.10 filter paper and concentrated at 40°C using a rotary evaporator. The metabolic profiling of SPO was analysed by GC-MS technique. The MIC, MBC, activity index, and total antimicrobial activity of SPO were evaluated using standard procedures. Anti-inflammatory activity of SPO was screened using Carrageenan induced paw oedema and Cotton pellet-induced granuloma. Tail immersion test, Acetic acid writhing response and Formalin paw lick test was performed in rats in order to examine the analgesic activity of SPO. Wound healing activity of SPO was investigated by performing the incision wound model, Excision wound model and Dead space wound model in rats. RESULTS: The SPO displayed a constant degree of antimicrobial activity against B. cereus, B. subtilis, E. coli, A. niger, A. fumigatus and C. albicans with significant anti-inflammatory and analgesic activities. Also, a prominent wound healing potential of it was observed in excision, incision and dead space wound models with considerable elevation in granulation tissue hydroxyproline, hexuronic acid and hexosamine content in association with remarkable regulation of anti-inflammatory and antioxidant markers i.e., Lipid peroxidase (LPO), Nitric Oxide (NO), Superoxide dismutase (SOD), Glutathione (GSH), Catalase (CAT), Glutathione Peroxidase (GPx). CONCLUSION: These findings strongly validate the therapeutic potential of S. pubescens fruit essential oil in antimicrobial and anti-inflammatory mediated wound healing and suggests its promising application as valuable and novel indigenous leads in the food and pharmaceutical industries. To the best of our knowledge, this is the first-ever investigatory report on the systematic phytochemical and therapeutic examination of S. pubescens fruit essential oil.

Anti-Methanogenic Traits of Safflower Oil Compounds Against Methyl-Coenzyme M Reductase Receptor in Equines: An In Silico Docking Analysis.

Greenhouse gases emission from livestock is the major concern for the ecosystem. Despite the lower contribution of non-ruminants towards greenhouse gas emission as compared to the ruminants, the emission of methane (CH4) gas from equines is expected to be increased in future due to its increasing population. Thus, it is essential to find or screen potential anti-methanogenic agent in a cost-effective and quicker manner. Considering this, the present investigation was aimed to analyze anti-methanogenic characteristic of bioactive compounds of safflower oil by targeting methanogenesis catalyzing enzyme (Methyl-coenzyme M reductase; MCR) via in silico tool. Initially, a total of 25 compounds associated with safflower oil were selected and their drug-likeness traits were predicted through Lipinski's rule of 5. Of 25 compounds, 9 compounds passed all the parameters of Lipinski's rule of five. These 9 ligands were further submitted for ADME traits analysis using Swiss ADME tool. Results revealed the absence of Lipinski's violation and approval of drug-likeness attributes of methyl tetradecanoate, 3-isopropyl-6-methylenecyclohex-1-ene, trans-2,4-decadienal, cis-6-nonenal, limonene, syringic acids, matairesinol, acacetin, and 2,5-octanedione. Molecular docking analysis was performed for analyzing the affinity between the selected 9 ligands and MCR receptor using FRED v3.2.0 from OpenEye Scientific Software and Discovery Studio client v16.1.0. Results showed maximum binding interaction of acacetin with MCR with the chemguass4 score of -13.35. Other ligands showed comparatively lower binding affinity in the order of matairesinol (-12.43) > methyl tetradecanoate (-9.25) > cis-6-nonenal (-7.88) > syringic acids (-7.73) > limonene (-7.18) > trans-2,4-decadienal (-7.07) > 3-isopropyl-6-methylenecyclohex-1-ene (-7.01) > 2,5-octanedione (-7.0.). In a nutshell, these identified compounds were observed as potential agents to reduce CH4 production from equines by targeting MCR. This in silico study emphasized the role of safflower-associated compounds in developing anti-methanogenic drug for equines in future.

Hydroxyl containing benzo[b]thiophene analogs mitigates the acrylamide induced oxidative stress in the zebrafish larvae by stabilizing the glutathione redox cycle.

AIMS: This study aims to elucidate a systematic free-radical quenching ability of synthesized benzo[b]thiophene derivatives using in vitro assays and acrylamide induced oxidatively stressed model in zebrafish larvae. MATERIALS AND METHODS: Antioxidant activity of the compounds was evaluated using in vitro methods. The toxicity of the compounds was evaluated in Madin-Darby Canine Kidney (MDCK) cell line and zebrafish embryos. Oxidative stress was generated by acrylamide (1 mM) in zebrafish larvae and treated with compounds to evaluate the in vivo antioxidant ability. Specific fluorescence dyes were used to detect ROS generation, lipid peroxidation, and cell death followed by gene expression using RT PCR. Density functional theory (DFT) and in silico pharmacokinetics were also studied. KEY FINDINGS: Compound BP and EP have a greater in vitro free radical scavenging ability. The maximum tolerated concentration (MTC) of the compounds in zebrafish larvae is 80 µM. The antioxidant system in zebrafish larvae was dysregulated due to acrylamide exposure and improvement was found while treating acrylamide exposed larvae with compounds 1-(3-hydroxybenzo[b]thiophen-2-yl) ethanone (BP) and 1-(3-hydroxybenzo[b]thiophen-2-yl) propan-1-one hydrate (EP). Compound BP and EP enhanced the SOD and CAT activity, reduced the ROS and lipid peroxidation level, thus decreasing cell death in zebrafish larvae. Compound BP and EP also improved the glutathione redox cycle by stabilizing glutathione-related gene expressions. SIGNIFICANCE: Hydroxyl-containing compounds BP and EP are promising lead molecules for pathological conditions related to oxidative stress, which showed an attenuated effect on acrylamide-induced oxidative stress in zebrafish larvae by enhancing the glutathione redox cycle and enzymatic antioxidants.

Protective Effect of Borassus flabellifer Haustorium Extract against Alkoxyl Radical-Induced Cytotoxicity by Improving Glutathione Metabolism by Modulating Nrf2/Haeme Oxygenase-1 Expression.

AIM: The study was aimed to assess the ability of Borassus flabellifer haustorium methanolic extract (BHE) on de novo glutathione biosynthesis in normal and pro-oxidant exposed cells via Nuclear factor erythroid 2-related factor 2 (Nrf2) and haeme oxygenase-1 (HO1) signaling in 2,2'-Azobis(2-methylpropionamidine) di-hydrochloride (AAPH) induced cytotoxicity in normal intestinal epithelial cells (IEC-6 cells). METHODS: The in vitro antioxidant activity was determined in terms of radical scavenging and ex vivo hemolysis. The cytoprotective effect was studied using AAP H as the alkoxyl radical inducer in IEC-6 cell model. The mechanistic basis of protection is determined by Nrf2/HO1 expression using qPCR. RESULTS: In vitro screening observed DPPH, hydrogen peroxide and ABTS radical scavenging activity for the BHE; further, BHE also protected the oxidative hemolysis in the erythrocytes induced by AAPH. In IEC-6 cells, AAPH treatment significantly reduced the cell viability (p < 0.001) by inducing lipid peroxidation. Further, there observed a significant reduction in the activities of enzymes involved in the de novo glutathione biosynthesis (p < 0.01) and glutathione reductase in these cells. However, pretreatment with BHE (10, 25 and 50 µg/mL) dose-dependently protected from the cytotoxicity of AAPH-derived alkoxyl radicals (p < 0.05); besides, the de novo glutathione biosynthesis and regeneration of GSH from oxidized form was also increased in these cells. In corroboration with the biochemical parameters, the Nrf2/HO1 expression was upregulated by the BHE pretreatment concomitantly reducing the cellular lipid peroxidation products. The improvement glutathione biosynthesis was also observed in BHE alone treated cells. CONCLUSION: The study indicated the potential of methanolic extract of Borassus flabellifer haustorium in enhancing the de novo glutathione biosynthesis in normal and pro-oxidant exposed cells by Nrf2/HO1 dependent manner, concomitantly mitigating the toxicity of AAPH-derived alkoxyl radicals in intestinal epithelial cells.

Borassus flabellifer Linn haustorium methanol extract mitigates fluoride-induced apoptosis by enhancing Nrf2/Haeme oxygenase 1 -dependent glutathione metabolism in intestinal epithelial cells.

Fluoride is the most common cause of drinking water-associated toxicity and is known to induce various metabolic imbalances and dental/skeletal fluorosis. The present study analyzed the protective effect of Borassus flabellifer Linn. haustorium extract (BHE) against fluoride-induced intestinal redox metabolism and apoptosis. The total polyphenols and total flavonoids present in BHE were estimated to be 39.67 ± 5.14 mg gallic acid equivalent/g extract and 8.59 ± 0.74 mg quercetin equivalent. In cultured intestinal epithelial cells (IEC-6), sodium fluoride exposure-induced apoptosis mediated through antioxidant enzyme inhibition and subsequent oxidative damages. Further, there observed an increased expression of caspase-3, caspase-7, and apoptotic protease activating factor-1 (apaf-1) genes, increased cytochrome C release, and caspase 3/7 activity indicating the apoptosis- mediated cell death (p < 0.05). Upon pretreatment with BHE, the cytotoxic effect of fluoride was reduced by decreasing the expression of apoptotic genes and increased the cytochrome release as well as caspase 3/7 activity (p < 0.01). Providing the mechanistic basis, the expression of nuclear factor erythroid 2-related factor-2 (Nrf2)/haeme oxygenase-1 (HO1) gene was increased in the BHE pretreated cells; corroborating to these, there observed increased activity of glutathione biosynthetic enzymes (p < 0.05) and glutathione reductase. Hence, the protective effect of BHE may be mediated through Nrf2-mediated glutathione biosynthesis, the subsequent establishment of redox balance, and inhibition of apoptosis in intestinal epithelial cells.

Analysis of the chemical composition of root essential oil from Indian sarsaparilla (Hemidesmus indicus) and its application as an ecofriendly insecticide and pharmacological agent.

The Indian sarsaparilla (Hemidesmus indicus) is a commonly used plant in Indian traditional medicine of Ayurveda for the preparation of various non-alcoholic beverages. However, limited studies are available on the essential oil of H. indicus roots (HRO); therefore, the study evaluated the antioxidant, anti-inflammatory and antidiabetic activities of H. indicus root essential oil as well as insecticide potential against the common pests of stored food materials (Sitophilus oryzae, Callosobruchus maculatus and Tribolium castaneum). The repellant efficacy of HRO was found to be high against S. oryzae (8.21 ± 0.55 µg/mL). Likewise, the fumigant potential was also observed for HRO against these pests; the higher activities were observed against S. oryzae and C. maculatus (32.46 ± 1.42 and 35.18 ± 1.62 µg/L). Besides, the essential oil was also found to be active as a contact poison, however, against all the three pests, the toxicity was above 100 µg/mm3, being the highest against C. maculatus (122.8 ± 3.57 µg/mm3). To analyze the possible effect of the essential oil on grains, the different grains were allowed to germinate and compared to that of normal; thus, the non-toxic nature of HRO against the stored products is also confirmed. The essential oil shown to have DPPH hydrogen peroxide and ABTS radical scavenging, nitric oxide scavenging potential, and inhibition of lipoxgenase, alpha-amylase and alpha-glucosidase. Overall, the present study concludes that the H. indicus may be a suitable repellant and fumigant agent against different pests of stored products and a possible antioxidant, anti-inflammatory, and anti-diabetic agent.

Phytochemical profiling of Azima tetracantha Lam. leaf methanol extract and elucidation of its potential as a chain-breaking antioxidant, anti-inflammatory and anti-proliferative agent.

Azima tetracantha, a traditional medicinal plant included in the order Brassicales and family Salvadoraceae, is widely used as a dietary supplement in folklore medicines. The plant is also used for the treatment of rheumatism, diarrhea and other inflammatory disorders. The present investigation focused on the phytochemical composition, radical scavenging, reducing potential and anti-proliferative activities of the A. tetracantha leaves. Quantitative estimation of the polyphenols and flavonoids revealed significantly elevated levels in the methanol extract. Corroborating with this, methanol extract exhibited higher in vitro anti-radical scavenging effect against 2,2-diphenyl-1- picrylhydrazyl (34.14 ± 2.19 µg/mL), and hydrogen peroxide (44.96 ± 1.77 µg/mL), as well as ferric reducing properties (58.24 ± 6.98 µg/mL). The methanolic extract also showed strong lipoxygenase (71.42 ± 6.36 µg/mL) and nitric oxide inhibitory activities (94.23 ± 8.11 µg/mL). Cytotoxic activity against MCF7 cells was found to be higher (IC50= 37.62 ± 2.94 µg/mL), than that of MDAMB231 cells (IC50= 69.11 ± 5.02 µg/mL). The qPCR-based analysis indicated dose-dependent increase in the expression of the pro-apoptotic genes such as executioner caspases and apoptotic protease activating factor-1. Overall, the results indicated the possible use of methanol extract of A. tetracantha leaves as a chain-breaking antioxidant molecule and are capable of inhibiting inflammatory enzymes and the proliferative potential of breast cancer cells.

Chemical Composition, Insecticidal and Mosquito Larvicidal Activities of Allspice (Pimenta dioica) Essential Oil.

Essential oils are biologically and environmentally safe pesticidal compounds yielded from aromatic plants. Spices are important sources of essential oils, and they are widely used in the medicine, food, and various other industries. Among the different spices, Allspice (Pimenta dioica) is underexplored in terms of its biological efficacy and a limited number of studies are available on the chemical composition of Allspice essential oil (AEO); thus, the present study evaluated the larvicidal property, the repellency, and the fumigant toxicity against common pests of stored products of AEO. AEO was found to inhibit the survival of larvae of such vectors as Aedis, Culex, and Armigeres species. Further, AEO was found to exert repellant effects against the pests of such stored products as Sitophilus, Callosobruchus, and Tribolium. Similarly, the fumigant toxicity was found to be high for AEO against these species. The contact toxicity of AEO was high against Sitophilus and Callosobruchus. Apart from that, the essential oil was found to be safe against a non-target organism (guppy fishes) and was found to be non-genotoxic in an Allium cepa model. Overall, the results of the present study indicate that the essential oil from Allspice could be used as an environmentally safe larvicidal and biopesticidal compound.

Antimicrobial and anti-biofilm activities of polyphenols extracted from different Saudi Arabian date cultivars against human pathogens.

BACKGROUND: Microbial diseases have emerged as a serious threat to the healthcare system globally and also in Saudi Arabia; various diseases are associated with higher mortality and increased spreading rates. Dietary sources are often entertained to improve the immune system of the body to fight against these infections. The date fruit (Phoenix dactylifera L) is one such functional food that is less explored for its actual potentials. METHODS: The polyphenols isolated from the different cultivars of dates in Saudi Arabia (Ajwa, Safawi, Khalas, and Sukkary) was evaluated for their antibacterial, anti-fungal and anti-biofilm forming abilities. The anti-radical properties of the phenolic extract were conducted in terms of the scavenging of diphenyl-1-picrylhydrazyl radical, hydrogen peroxide radicals and ABTS radicals. The anti-inflammatory potential was analyzed in terms of lipoxygenase activity inhibition. RESULTS: The phenolic compounds of different dates were spectrophotometrically estimated and cultivars such as Ajwa and Sukkary had the highest polyphenol content. It was also noteworthy that they exerted potent antibacterial activities against Staphylococcus aureus, Streptococcus pyogenes, and Pseudomonas aeruginosa as estimated by the least minimal inhibitory concentration (MIC) or minimal bactericidal concentration (MBC) values. Besides, these polyphenols reduced the biofilm formation by these bacterial pathogens; it is thus possible that the polyphenols from dates can be a promising antimicrobial agent against various pathogenic strains. Corroborating with these, the polyphenol extracts from different Saudi Arabian dates were found to inhibit the growth of mycelium in strains of Candida albicans and Aspergillus niger. Apart from these, the polyphenol isolates also exhibited significant anti-radical properties against different reactive radical systems. CONCLUSION: Overall, the efficacy of phenolic compounds extracted from the dried date fruits are not only restricted to the functional food features; instead, these molecules are capable of preventing the growth of microbial pathogens of humans. Hence, it may emerge as potent antibacterial, anti-fungal and anti-biofilm forming candidates.

Toxic effects of fluoride in intestinal epithelial cells and the mitigating effect of methanol extract of coconut haustorium by enhancing de novo glutathione biosynthesis.

Fluoride ions are an important environmental contaminant and pollutant found in a wide variety of environmental conditions. The fluoride in drinking water is evident to induce toxic effects including neurodegeneration, skeletal and dental fluorosis as well as organ damage. Nutraceuticals and functional foods are emerging as possible preventive agents against fluoride toxicity. Hence, the possible use of an emerging functional food-the coconut haustorium is being evaluated against sodium fluoride-induced toxicity in intestinal cells (IEC-6). The cells exposed to fluoride showed significant cell death mediated through the increased lipid peroxidation and glutathione depletion. The glutathione biosynthetic enzymes were inhibited by the exposure to fluoride and the apoptotic genes (caspases 3/7 and apaf-1) were upregulated. The CHE pre-treatment improved the activity of enzymes involved in the de novo biosynthesis of glutathione and subsequently improved the intracellular GSH pool. The improved antioxidant defense was also evident from the reduced expression of apoptotic genes (p < 0.05). Overall, the study concludes that fluoride ions induce oxidative stress-mediated apoptosis in intestinal epithelial cells, via inhibiting glutathione biosynthesis. Methanol extract of coconut haustorium increased glutathione biosynthesis and subsequently prevented fluoride toxicity in IEC-6 cells by virtue of its antioxidant potentials.