GC-MS Analysis and In-vitro Apoptosis Induction and Anticancer Activity of Methanol Extract of Aspergillus terreusagainst Lung Cancer

The present study was focusing on qualitative and quantification of bioactive compounds present in Aspergillus terreus and evaluating its anticancer activity and apoptosis detection against lung cancer. Methods: A. terreus was sequentially extracted using the Soxhlet extraction technique with hexane, ethyl acetate, methanol and distilled water. Detection of bioactive compounds was done using Standard biochemical tests and GC-MS analysis was performed with NIST database to identify the bioactive compounds. The toxicity and anticancer activity of crude extract was investigated using MTT assay on L929 cells and lung cancer A549 cells whereas apoptosis study was conducted through Flowcytometry-based surface marker study on the A549 cancer cell line. Results: secondary metabolites analysis showed the presence of phenols and terpenoids as major constituents in the methanol extract whereas other solvent extracts have shown the absence of major bioactive compounds. Quantification studies showed that methanol extract has shown the phenolic content 179 µg/g of Gallic acid equivalent. The GC-MS analysis showed the presence of 1-Flurodecane, Methyl palmitate, Ethyl palmitate, 9, 12-Octadecanopic acid, 10-Octadecanoic acid, Methyl stearate, Octadecadeoinoate, Ethyl 9-hexadecanoate and 1-Monoarachidin as major bioactive compounds. Further, MTT based toxicity study on the L929 cell line revealed that methanol extract at lower concentrations like 50µg, 100µg and150µg shown more than 50% of cell viability and at higher concentration between 200µg-250 µg it was showing toxic nature with 47.89±0.01% viability. In case of anticancer activity against lung cancer A549 cell line the methanol extract have shown the dose dependent activity i.e the percentage of cell viability was decreased with increase in the concentration of methanol extract at 250µg the cell viability was found to be 35.12±0.005%. Flow cytometry based apoptosis study revealed that methanol extract has shown the inducing apoptosis in treated lung cancer A549 cells with percentage of 10.84. Conclusion: overall the present study shown that A. terreus possess different class of bioactive compounds and it has higher phenolic content. Toxicity study showed that methanol extract exhibited toxic nature at higher concentration on tested cell line and Anticancer and Apoptosis study revealed that methanol extract has shown the prominent with inhibiting the growth of lung cancer A549 cells through inducing apoptosis. Further, A. terreus would be a promising natural microorganism that has to be further researched in order to discover and isolate potent drug to treat cancer. Future studies will be on study of in-vivo animal studies and study of molecular mechanism of drug action on particular with anticancer study

Synthesis, in vitro biological evaluation and molecular docking study of coumarin-1,4-dihydropyridine derivatives as potent anti-inflammatory agents

The green chemistry approach provides for the synthesis of coumarin-1,4-dihydropyridine scaffolds 6a-o via sequential multicomponent reaction using catalytic amount of triethylamine (TEA). These new coumarin scaffolds have been successfully explored for the effective inflammatory as well as microbial infection inhibitors. The antimicrobial activity results of the title compounds have shown potent activity against both gram positive and gram negative bacterial, and fungal stains. Additionally, anti-inflammatory activity of all the compounds has been found to be quite promising in comparison with standard Diclofenac sodium. Furthermore, the in silico docking study has been performed for all the compounds with S. aureus DNA gyrase and cyclooxygenase-2 (PDB ID 4PH9). The computational results are in good agreement with the in vitro antibacterial and anti-inflammatory experimental results.

Synthesis, in vitro biological evaluation and molecular docking study of coumarin-1,4-dihydropyridine derivatives as potent anti-inflammatory agents

418-432The green chemistry approach provides for the synthesis of coumarin-1,4-dihydropyridine scaffolds 6a-o via sequential multicomponent reaction using catalytic amount of triethylamine (TEA). These new coumarin scaffolds have been successfully explored for the effective inflammatory as well as microbial infection inhibitors. The antimicrobial activity results of the title compounds have shown potent activity against both gram positive and gram negative bacterial, and fungal stains. Additionally, anti-inflammatory activity of all the compounds has been found to be quite promising in comparison with standard Diclofenac sodium. Furthermore, the in silico docking study has been performed for all the compounds with S. aureus DNA gyrase and cyclooxygenase-2 (PDB ID 4PH9). The computational results are in good agreement with the in vitro antibacterial and anti-inflammatory experimental results

Physicochemical properties, antioxidant and anti-inflammatory activities of coumarin-carbonodithioate hybrids

Objective: To study physicochemical properties, antioxidant and anti-inflammatory activities of coumarin-carbonodithioate hybrids. Methods: The substituted 4-bromomethyl coumarins were synthesized in first step by the cyclization. Then the reaction of substituted coumarins (a-e) with potassium O-ethyl/methyl carbonodithioate (1) by using absolute ethanol as solvent, afforded coumarin-carbonodithioate (1a-1j) derivatives under microwave irradiation and the conventional method. The spectroscopic analysis was used for the characterization of coumarin derivatives. The title (1a-1j) compounds were confirmed by spectroscopic methods. Antioxidant property was evaluated by using DPPH free radical-scavenging ability assay method and anti-inflammatory activity was evaluated by protein denaturation procedure using diclofenac sodium as a standard. Drug-likeness. In-silico toxicity was predicted with LD50 value and bioactivity score was also calculated for all the compounds. Results: All coumarin (1a-1j) compounds exhibited promising in-vitro antioxidant and anti-inflammatory properties in comparison to standard drugs. All tested compounds were used for evaluating their physicochemical properties as set by Lipinski rule. It was observed that the synthesized compounds followed rule of five, indicating more ‘drug-like’ nature. Conclusions: All the screened coumarin-carbonodithioates display promising in vitro antioxidant and anti-inflammatory activities. From the physicochemical properties of coumarin derivatives, it is found that none of the compounds violate the Lipinski rule and they fall well in the range of rule of five. It is concluded that the coumarin-carbonodithioate hybrids act with more ‘drug-like’ nature

A pilot study: Changes of MDAMB-231 cancer cell line response to synthesized oleic acid – coated MgFe2O4 nano ferrite compound and its cytotoxic effects on L929 cell line

In this investigation, we synthesized MgFe2O4 nano ferrite compounds using the sol-gel method and coated them with oleic acid. The synthesized compounds were evaluated for cytotoxicity study on the L929 cell line and the anticancer activity against the breastcancer-MDAMB-231 cellline using in vitro approach. The physical structure of the compounds was confirmed to have a cubic spinel structure and Fd3m space group using X-ray diffraction (XRD). Scherrer's formula revealed crystallite sizes of 11 nm for MgFe2O4and 33 nm for oleic acid-coated MgFe2O4 nanoferrites. Fourier transform infrared (FTIR) spectroscopy confirmed the existence of MgFe2O4 inuncoated and coated compounds. Scanning electron microscopy (SEM) was used to examine the surface structure and morphology of the synthesized compounds. The compounds show a grain-like structure with a little agglomeration. With the help of histogram graph average size of the particles are determined, where the MgFe2O4 and coated MgFe2O4 have 17.2 and 20 nm average particle sizes. Testing of magnetic properties was done with a vibrating sample magnetometer (VSM), demonstrating ferromagnetic properties for the samples. Cytotoxicity tests revealed that MgFe2O4 exhibited higher cytotoxicity compared to oleic acid-coated MgFe2O4 nanoferrite on L929 cells. MgFe2O4 & oleic acid-coated MgFe2O4 compounds were tested on the MDAMB-231 breast cancer cell line, with the uncoated sample displaying significantly greater anticancer activity, as evidenced by a higher percentage of cell viability and a lower IC50 value of 55.67 µg compared to 65.97 µg for the coated sample

Green Synthesis and Characterization of Iron Nanoparticles Synthesized from Aqueous Leaf Extract of Vitex leucoxylon and Its Biomedical Applications

The cold extraction method was used to obtain the aqueous extract of Vitex leucoxylon leaves in a ratio of 1:10. Iron nanoparticles (FeNPs) were synthesized using aqueous leaf extract of V. leucoxylon as a reducing agent. The phytoreducing approach was used to make FeNPs by mixing 1 mL of plant extract with 1 mM of ferric sulfate. Scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), Ultraviolet–visible spectroscopy (UV-Vis), and energy-dispersive X-ray spectroscopy were used to examine the synthesized FeNPs. The reducing reaction was shown by a change in the color of the solution, and the formation of black color confirms that FeNPs have been formed. The greatest absorption peak (max) was found at 395 nm in UV-Vis spectral analysis. The FTIR spectra of V. leucoxylon aqueous leaf extract showed shifts in some peaks, namely 923.96 cm−1 and 1709.89 cm−1, with functional groups carboxylic acids, unsaturated aldehydes, and ketones, which were lacking in the FTIR spectra of FeNPs and are responsible for FeNPs formation. FeNPs with diameters between 45 and 100 nm were observed in SEM images. The creation of FeNPs was confirmed by EDX, which shows a strong signal in the metallic iron region at 6–8 Kev. XRD revealed a crystalline nature and an average diameter of 136.43 nm. Antioxidant, anti-inflammatory, cytotoxic, and wound healing in vitro tests reported significant activity of the FeNPs. The cumulative findings of the present study indicate that the green synthesis of FeNPs boosts its biological activity and may serve as a possible dermal wound-healing agent and cytotoxic agent against cancer. Future study is needed on the identification of mechanisms involved in the synthesis of FeNPs by V. leucoxylon and its biomedical applications

Synthesis and Characterization of Silver Nanoparticles from Rhizophora apiculata and Studies on Their Wound Healing, Antioxidant, Anti-Inflammatory, and Cytotoxic Activity

Silver nanoparticles (AgNPs) have recently gained interest in the medical field because of their biological features. The present study aimed at screening Rhizophora apiculata secondary metabolites, quantifying their flavonoids and total phenolics content, green synthesis and characterization of R. apiculata silver nanoparticles. In addition, an assessment of in vitro cytotoxic, antioxidant, anti-inflammatory and wound healing activity of R. apiculata and its synthesized AgNPs was carried out. The powdered plant material (leaves) was subjected to Soxhlet extraction to obtain R. apiculata aqueous extract. The R. apiculata extract was used as a reducing agent in synthesizing AgNPs from silver nitrate. The synthesized AgNPs were characterized by UV-Vis, SEM-EDX, XRD, FTIR, particle size analyzer and zeta potential. Further aqueous leaf extract of R. apiculata and AgNPs was subjected for in vitro antioxidant, anti-inflammatory, wound healing and cytotoxic activity against A375 (Skin cancer), A549 (Lung cancer), and KB-3-1 (Oral cancer) cell lines. All experiments were repeated three times (n = 3), and the results were given as the mean ± SEM. The flavonoids and total phenolics content in R. apiculata extract were 44.18 ± 0.086 mg/g of quercetin and 53.24 ± 0.028 mg/g of gallic acid, respectively. SEM analysis revealed R. apiculata AgNPs with diameters ranging from 35 to 100 nm. XRD confirmed that the synthesized silver nanoparticles were crystalline in nature. The cytotoxicity cell viability assay revealed that the AgNPs were less toxic (IC50 105.5 µg/mL) compared to the R. apiculata extract (IC50 47.47 µg/mL) against the non-cancerous fibroblast L929 cell line. Antioxidant, anti-inflammatory, and cytotoxicity tests revealed that AgNPs had significantly more activity than the plant extract. The AgNPs inhibited protein denaturation by a mean percentage of 71.65%, which was equivalent to the standard anti-inflammatory medication diclofenac (94.24%). The AgNPs showed considerable cytotoxic effect, and the percentage of cell viability against skin cancer, lung cancer, and oral cancer cell lines was 31.84%, 56.09% and 22.59%, respectively. R. apiculata AgNPs demonstrated stronger cell migration and percentage of wound closure (82.79%) compared to the plant extract (75.23%). The overall results revealed that R. apiculata AgNPs exhibited potential antioxidant, anti-inflammatory, wound healing, and cytotoxic properties. In future, R. apiculata should be further explored to unmask its therapeutic potential and the mechanistic pathways of AgNPs should be studied in detail in in vivo animal models

<i>Tribulus terrestris</i> Cytotoxicity against Breast Cancer MCF-7 and Lung Cancer A549 Cell Lines Is Mediated via Activation of Apoptosis, Caspase-3, DNA Degradation, and Suppressing Bcl-2 Activity

The primary objective of this research was to use flow cytometry to gain mechanistic insights into the cytotoxic effects of Tribulus terrestris extracts on breast cancer (MCF7) and lung cancer (A549) cell lines. T. terrestris was extracted using a Soxhlet apparatus in a progressive process. GC–MS was used to establish the phytochemical constituents. The amounts of phenolic compounds and flavonoids in the plant extracts were calculated using spectrophotometric analysis. The cytotoxicity of plant extracts was initially evaluated in non-malignant L929 cells, then in carcinogenic MCF-7 and A549 cell lines. Then, we performed an Annexin V assay, an anti-Bcl-2 assay, a Caspase-3 assay, and a DNA fragmentation (TUNEL) assay, using flow cytometry to investigate the underlying molecular processes. Based on the data, the methanolic extract of T. terrestris contained the highest amounts of phenolic compounds and flavonoids, with values of 169.87 µg GAE/g dwt and 160.12 µg QE/g dwt, respectively. Analysis by GC–MS revealed the presence of bioactive phytochemicals with proven cytotoxicity. Based on the MTT experiment, we determined that the IC50 values for the methanol extract’s effect on the viability of the MCF-7 and A549 cell lines were 218.19 and 179.62 µg/mL, respectively. The aqueous and methanol extracts were less cytotoxic when tested against the cancer-free L929 cell line (IC50 = 224.35 µg/mL). In both breast and lung cancer cells, the methanolic extract was found to activate caspase-3 and inhibit the Bcl-2 protein, resulting in early and late apoptosis and cell death via DNA damage. These findings point to cytotoxic effects of T. terrestris methanol extract against breast and lung cancer cell lines. Due to its potential as a source of anti-cancer chemotherapeutic medicines, T. terrestris warrants further investigation

In Vitro Cytotoxicity and Spectral Analysis-Based Phytochemical Profiling of Methanol Extract of <i>Barleria hochstetteri,</i> and Molecular Mechanisms Underlying Its Apoptosis-Inducing Effect on Breast and Lung Cancer Cell Lines

The objectives of this research were to carry out GC–MS and LC–MS-based phytochemical profiling of Barleria hochstetteri, as well as flow cytometry-based mechanistic investigations of the cytotoxic effect of its extracts against breast and lung cancer cell lines. This preclinical in vitro study was carried out in Saudi Arabia and India, from 11 August to 15 January 2022. Barleria hochstetteri was sequentially extracted using the Soxhlet extraction technique. Utilizing LC–MS and GC–MS methods, the phytochemical profiling was performed. Additionally, the total phenolic compounds and flavonoids were quantified in the plant extract using spectrophotometric techniques. In this study, we first examined the cytotoxicity of the plant extract on non-malignant L929 cells and on the carcinogenic MCF-7 and A549 cell lines. Then, we studied the underlying molecular pathways by means of Anti-Bcl-2, caspase-3, and DNA fragmentation (TUNEL) assays, using flow cytometry. The results revealed phenolic compounds and flavonoids to be the two major components in the methanolic extract of B. hochstetteri, with concentrations of 3210 µg GAE/g dwt and 1863 µg QE/g dwt, respectively. Results from GC–MS and LC–MS analyses revealed the presence of bioactive phytochemicals with known cytotoxicity. From the MTT assay on cell viability, the IC50 of the methanol extract for the MCF-7 and A549 cell lines were 219.67 and 144.30 µg/mL, respectively. With IC50 values of 324.24 and 266.66 µg/mL, respectively, the aqueous and methanol extracts were less toxic when tested against the non-cancerous L929 cell line. The extract caused early and late apoptosis in the tested breast and lung cancer cells by activating caspase-3 and inhibiting Bcl-2 protein, and it also caused cell death via DNA damage, based on flow cytometric and molecular marker analyses. These findings indicate that the methanol extract of B. hochstetteri was cytotoxic on breast cancer and lung cancer cell lines. To uncover cancer-fighting chemicals, there is a need for further research on B. hochstetteri, as it is a promising source of anti-cancer chemotherapeutic drugs

Apoptotic Cell Death via Activation of DNA Degradation, Caspase-3 Activity, and Suppression of Bcl-2 Activity: An Evidence-Based Citrullus colocynthis Cytotoxicity Mechanism toward MCF-7 and A549 Cancer Cell Lines

The objectives of this study are to investigate the cytotoxic effect of different Citrullus colocynthis extracts on breast and lung cancer cell lines using flow cytometry to gain mechanistic insights. C. colocynthis was extracted sequentially using the Soxhlet method. We first tested the plant extracts&rsquo; cytotoxicity on non-malignant L929 cells and cancerous breast (MCF-7) and lung (A549) cell lines. We observed that the IC50 of the methanol extract on the viability of MCF-7 and A549 cell lines was 81.08 &micro;g/mL and 17.84 &micro;g/mL, respectively, using the MTT assay. The aqueous and methanol extracts were less toxic when tested against the non-cancerous L929 cell line, with IC50 values of 235.48 &micro;g/mL and 222.29 &micro;g/mL, respectively. Then, using flow cytometry, we investigated the underlying molecular pathways with Annexin-V, Anti-Bcl-2, Caspase-3, and DNA fragmentation (TUNEL) assays. Flow cytometric and molecular marker analyses revealed that the methanol extract activated caspase-3 and inhibited Bcl-2 protein, causing early and late apoptosis, as well as cell death via DNA damage in breast and lung cancer cells. These findings indicate that the methanol extract of C. colocynthis is cytotoxic to breast and lung cancer cell lines. The total phenolic and flavonoid content analysis results showed the methanolic extract of C. colocynthis has a concentration of 326.25 &mu;g GAE/g dwt and 274.61 &mu;g QE/g dwt, respectively. GC-MS analysis of the methanol extract revealed phytochemicals relevant to its cytotoxicity