Impurity-profiling UPLC methods for quantitative analysis of some antiemetics formulated with pyridoxine.

Cyclizine hydrochloride (CYC) and meclozine hydrochloride (MEC) are antihistaminic drugs generally co-formulated with pyridoxine hydrochloride (PYR) to treat nausea and vomiting in pregnancy. Several analytical techniques have been applied for the determination of CYC or MEC with PYR, but determination of CYC impurity; benzhydrol (BEH) or MEC impurity; or 4-chlorobenzophenone (BEP) has not been paid attention to. Therefore, micellar UPLC method is introduced for analysis of ternary mixtures containing PYR together with both CYC and BEH (mixture I) or MEC and BEP (mixture II). Chromatographic separation was achieved using a Hypersil gold C8 column (50 × 2.1 mm, 1.9 µm) using 0.01 M sodium dodecyl sulfate modified to pH 3.5 using phosphoric acid:acetonitrile (45:55 by volume) for mixture I and 0.1% sodium dodecyl sulfate, 0.1% sodium bicarbonate adjusted to pH 2.6 by phosphoric acid:acetonitrile (47:53 by volume) for mixture II as mobile phases. The separated peaks were detected at 230 and 245 nm for mixtures I and II, respectively. The adopted methods were validated in conformance with the International Conference on Harmonization (ICH) recommendations and were properly applied in commercial pharmaceutical formulation analysis. Comprehensive ecological comparison was achieved, confirming a higher ecological value of the presented methods compared to the earlier reported methods.

Highly sensitive detection of kojic acid in food samples using fluorescent carbon dots derived from pomegranate peel.

Kojic acid (KA) has gained significant attention due to its widespread use in the food and cosmetics industries. However, concerns about its potential carcinogenic effects have heightened the need for sensitive detection methods. This study introduces a fluorescence-based optical sensor for the quantification of KA in food samples, utilizing fluorescent carbon dots (CDs) synthesized from pomegranate peel via a hydrothermal method. The Stern-Volmer plot demonstrated a linear response for KA in the range of 120 to 1200 µM, with a Pearson correlation coefficient (r) of 0.9999 and. The sensor exhibited a detection limit of 30 ± 0.04 µM and a limit of quantification (LOQ) of 90 ± 0.14 µM. Application of the developed method to soy sauce and vinegar samples yielded accurate KA determinations, with recoveries of 103.11 ± 0.96% and 104.45 ± 2.15%, respectively. These findings highlight the potential of the proposed sensor for practical applications in food quality and safety assessment, offering valuable insights into the presence of KA in food products.

Quality-by-design ecofriendly potentiometric sensor for rapid monitoring of hydroxychloroquine purity in the presence of toxic impurities.

Hydroxychloroquine (HCQ) is prescribed to treat malaria and certain autoimmune diseases. Recent studies questioned its efficiency in relieving COVID-19 symptoms and improving clinical outcomes. This work presents a quality-by-design approach to develop, optimize, and validate a potentiometric sensor for the selective analysis of HCQ in the presence of its toxic impurities (key starting materials), namely 4,7-Dichloroquinoline (DCQ) and hydroxynovaldiamine (HND). The study employed a custom experimental design of 16 sensors with different ion exchangers, plasticizers, and ionophores. We observed the Nernstian slopes, correlation coefficients, quantification limit, response time, and selectivity coefficient for DCQ and HND. The computer software constructed a prediction model for each response. The predicted responses strongly correlate to the experimental ones, indicating model fitness. The optimized sensor achieved 93.8% desirability. It proved a slope of 30.57 mV/decade, a correlation coefficient of 0.9931, a quantification limit of 1.07 × 10-6 M, a detection limit of 2.18 × 10-7 M, and a fast response of 6.5 s within the pH range of 2.5-8.5. The sensor was successfully used to determine HCQ purity in its raw material. The sensor represents a potential tool for rapid, sensitive, and selective monitoring of HCQ purity during industrial production from its starting materials.

A chemically modified solid-state sensor for magnesium(ii) ions and esomeprazole magnesium potentiometric assay.

The use of electrochemical sensors offers a simple, affordable solution with great reliability. Magnesium is a mineral that the body requires to function properly. It encourages preserving a stable pulse, strong bones, and healthy blood pressure. Herein, a novel ion-selective electrode using esomeprazole magnesium trihydrate as an ion-association complex was developed for magnesium(ii) ion determination in mineral water, drug substances, and pharmaceutical formulations. The electrode response was optimized in terms of plasticizer type, ion exchanger concentration, and membrane composition. To find the best sensor combination, the initial optimization research was performed using eight different sensors. A membrane containing 20% esomeprazole magnesium trihydrate, 36% carbon, and 44% o-Nitrophenyl Octyl Ether (NPOE) as a plasticizer yielded the best potentiometric response. The developed sensor demonstrated a Nernstian response with a slope of 29.93 ± 0.1 mV per decade in the concentration range of 1.41 × 10-5 mol L-1 to 1 × 10-2 mol L-1. Within a pH range of 5-8, it had a low detection limit of 4.13 × 10-6 mol L-1. When compared to the official method, there are no statistically significant differences.

Experimentally designed potentiometric sensor for green real-time and direct assay of hazardous bromate in bakery products.

Bakeries add extra potassium bromate to the dough to make homogeneous, elastic, fluffy bread. Bromate causes renal damage and cancer. FAO/WHO stated that bromate residues shouldn't be in baked products. A potentiometric sensor's membrane recipe was optimized for sensitive and selective bromate assay. We planned a custom experimental design of 21 sensors that included numerical and categorical factors (NPPE: PVC, matrix%, membrane thickness, and ionophore type). We defined sensor performance outcomes (Nernstian slope, quantification limit, correlation coefficient, response time and selectivity), and each sensor's outcome was determined. The computer software developed a predictive model for each outcome and the desirability function suggested the optimum sensor recipe. The sensor achieved a slope of -63.54 mV/decade and detection limit of 2 × 10-6 mol/L. The greenness profile was evaluated by the National Environmental Approach Index protocol. The developed sensor represents a reliable, fast, in-site tool for the assay of bromate in bakery products.

A sustainable approach for the degradation kinetics study and stability assay of the SARS-CoV-2 oral antiviral drug Molnupiravir.

Molnupiravir (MPV) is the first direct-acting oral antiviral drug that effectively decreases nasopharyngeal infections with SARS-CoV-2 virus. The stability of MPV was tested by subjecting the drug to various stress conditions. The drug is liable to oxidative, acidic, and alkaline degradation and showed significant stability against thermal degradation. Mass spectrometry identified the degradation products and guided suggestion of the degradation patterns. Interestingly, while inspecting the UV-absorption spectra, we observed no absorbance at 270 nm for the products of the three degradation pathways (c.f. intact MPV). Direct spectrophotometry seemed a solution that perfectly fit the purpose of the stability assay method in our case. It avoids sophisticated instrumentation and complex mathematical data manipulation. The method determined MPV accurately (100.32% ± 1.62) and selectively (99.49% ± 1.63) within the linear range of 1.50 × 10-5-4.0 × 10-4 M and down to a detection limit of 0.48 × 10-5 M. The proposed method is simple and does not require any preliminary separation or derivatization steps. The procedure proved its validity as per the ICH recommendations. The specificity was assessed in the presence of up to 90% degradation products. The study evaluated the greenness profile of the proposed analytical procedure using the National Environmental Methods Index (NEMI), the Analytical Eco-Scale, and the Green Analytical Procedure Index (GAPI). The three metrics unanimously agreed that the developed procedure results in a greener profile than the reported method. The method investigated the degradation reactions' kinetics and evaluated the reaction order, rate constant, and half-life time for each degradation process.

Simultaneous determination of retinoic acid and hydroquinone in skin ointment using spectrophotometric technique (ratio difference method).

An innovative spectrophotometric method was developed for simultaneous determination of compounds with interfering spectra in binary mixtures without previous separation, showing significant advantages over the conventional methods regarding minimal data manipulation and applicability. The proposed method was applied for the determination of retinoic acid and hydroquinone in laboratory-prepared mixtures with mean percentage recoveries 100.13 ± 0.31 and 99.99 ± 0.04, respectively, and in their pharmaceutical formulation with mean percentage recoveries 100.13 ± 0.86 and 100.07 ± 0.58, respectively. The method was validated according to USP guidelines and can be applied for routine quality control testing.

Secondary metabolites from Lantana camara L. flowers extract exhibit in vivo anti-urolithiatic activity in adult Wistar albino rats.

The present study aims at evaluating potential of the ethanol extracts of L. camara leaves (LE), flowers (FlE) and roots (RE) in the treatment of renal calculi and characterising the secondary metabolites in the active extract. The results revealed that the FlE had significantly reduced the levels of kidney parameters (calcium, creatinine, urea, and uric acid) against ethylene glycol (EG) injuries, and restored the activity of glutathione peroxidase (GPx), superoxide dismutase and lipid peroxide malondialdehyde to the normal level. In addition, FlE significantly attenuated iNOS tissue expression caused by EG. The results obtained in this study suggest the potential value of the L. camara L. flowers as an antiurolithiatic agent.

Experimentally designed chemometric models for the assay of toxic adulterants in turmeric powder.

Turmeric is an indispensable culinary spice in different cultures and a principal component in traditional remedies. Toxic metanil yellow (MY), acid orange 7 (AO) and lead chromate (LCM) are deliberately added to adulterate turmeric powder. This work compares the ability of multivariate chemometric models with those of artificial intelligent networks to enhance the selectivity of spectral data for the rapid assay of these three adulterants in turmeric powder. Using a custom experimental design, we provide a data-driven optimization for the sensitive parameters of the partial least squares model (PLS), artificial neural network (ANN) and genetic algorithm (GA). The optimized models are validated using sets of genuine turmeric samples from five different geographical regions spiked with standard adulterant concentrations. The optimized GA-PLS and GA-ANN models reduce the root mean square error of prediction by 18.4%, 31.1% and 55.3% and 25.0%, 69.9% and 88.4% for MY, AO and LCM, respectively.

Experimentally designed electrochemical sensor for therapeutic drug monitoring of Ondansetron co-administered with chemotherapeutic drugs.

The experimental design extracts valuable information about the main effects and interactions from the least number of experiments. The current work constructs a solid-state sensor for selective assay of Ondansetron (OND) in pharmaceutical dosage form and plasma samples. During optimization, the Design Expert® statistical package constructed a custom design of 15 sensors with different recipes. We fed the software with the experimentally observed performance parameters for each sensor (slope, LOQ, correlation coefficient, and selectivity coefficient for sodium ions). The computer software analyzed the results to construct a prediction model for each response. The desirability function was adjusted to optimize the Nernstian slope, minimize the LOQ and selectivity coefficients, and maximize the correlation coefficient (r). The practical responses of the optimized sensor were close to those predicted by the model (slope = 60.23 mV/decade slope, LOQ = 9.09 × 10-6 M, r = 0.999, sodium selectivity coefficient = 1.09 × 10-3). The sensor successfully recovered OND spiked to tablets and human plasma samples with mean percentage recoveries of 100.01 ± 1.082 and 98.26 ± 2.227, respectively. Results were statistically comparable to those obtained by the reference chromatographic method. The validated potentiometric method can be used for fast and direct therapeutic drug monitoring of OND co-administered with chemotherapeutic drugs in plasma samples.

Adoption of Advanced Chemometric Methods for Determination of Pyridoxine HCl, Cyclizine HCl, and Meclizine HCl in the Presence of Related Impurities: A Comparative Study.

BACKGROUND: Noising is an undesirable phenomenon accompanying the development of widely used chemometric models such as partial least square regression (PLSR) and support vector regression (SVR). OBJECTIVE: Optimizations of these chemometric models by applying orthogonal projection to latent structures (OPLS) as a preprocessing step which is characterized by canceling noise is the purpose of this research study. Additionally, a comprehensive comparative study between the developed methods was undertaken highlighting pros and cons. METHODS: OPLS was conducted with PLSR and SVR for quantitative determination of pyridoxine HCl, cyclizine HCl, and meclizine HCl in the presence of their related impurities. The training set was formed from 25 mixtures as there were five mixtures for each compound at each concentration level. Additionally, to check the validity and predictive ability of the developed chemometric models, independent test set mixtures were prepared by repeating the preparation of four mixtures of the training set plus preparation of another four independent mixtures. RESULTS: Upon application of the OPLS processing method, an upswing of the predictive abilities of PLSR and SVR was found. The root-mean-square error of prediction of the test set was the basic benchmark for comparison. CONCLUSION: The major finding from the conducted research is that processing with OPLS reinforces the ability of models to anticipate the future samples. HIGHLIGHTS: Novel optimizations of the widely used chemometric models; application of a comparative study between the suggested methods; application of OPLS preprocessing methods; quantitative determination of pyridoxine HCl, cyclizine HCl and meclizine HCl; checking the predictive power of developed chemometric models; analysis of active ingredients in their pharmaceutical dosage forms.

Dual PI3K/Akt Inhibitors Bearing Coumarin-Thiazolidine Pharmacophores as Potential Apoptosis Inducers in MCF-7 Cells.

Breast cancer is the most common malignancy worldwide; therefore, the development of new anticancer agents is essential for improved tumor control. By adopting the pharmacophore hybridization approach, two series of 7-hydroxyl-4-methylcoumarin hybridized with thiosemicarbazone (V-VI) and thiazolidin-4-one moieties (VII-VIII) were prepared. The in vitro anticancer activity was assessed against MCF-7 cells adopting the MTT assay. Nine compounds showed significant cytotoxicity. The most promising compound, VIIb, induced remarkable cytotoxicity (IC50 of 1.03 + 0.05 µM). Further investigations were conducted to explore its pro-apoptotic activity demonstrating S-phase cell cycle arrest. Apoptosis rates following VIIb treatment revealed a 5-fold and 100-fold increase in early and late apoptotic cells, correspondingly. Moreover, our results showed caspase-9 dependent apoptosis induction as manifested by an 8-fold increase in caspase-9 level following VIIb treatment. Mechanistically, VIIb was found to target the PI3K-α/Akt-1 axis, as evidenced by enzyme inhibition assay results reporting significant inhibition of examined enzymes. These findings were confirmed by Western blot results indicating the ability of VIIb to repress levels of Cyclin D1, p-PI3K, and p-Akt. Furthermore, docking studies showed that VIIb has a binding affinity with the PI3K binding site higher than the original ligands X6K. Our results suggest that VIIb has pharmacological potential as a promising anti-cancer compound by the inhibition of the PI3K/Akt axis.

The dynamic interplay between AMPK/NFκB signaling and NLRP3 is a new therapeutic target in inflammation: Emerging role of dapagliflozin in overcoming lipopolysaccharide-mediated lung injury.

Acute lung injury (ALI) is one the most common causes of morbidity and mortality in critically ill patients. In this study, we examined for first time the role of dapagliflozin (DPGZ) in lipopolysaccharide (LPS)-induced ALI in rats and determined the underlying molecular mechanisms by evaluating the effects of DPGZ on adenosine monophosphate kinase (AMPK), nuclear transcription factor kappa B, nucleotide-binding and oligomerization domain-like receptor 3 inflammasome activation. Treatment of acute lung injured rats with either low dose (5 mg/kg) or high dose (10 mg/kg) DPGZ significantly decreased oxidative stress by decreasing malondialdehyde and nitric oxide tissue levels with a significant increase in spectrophotometric measurements of superoxide dismutase, catalase, and reduced glutathione levels. DPGZ treatment resulted in a significant anti-inflammatory effect as indicated by suppression in myeloperoxidase activity, MCP-1, IL-1ß, IL-18, and TNF-α levels. DPGZ treatment also increased p-AMPK/t-AMPK with a significant reduction in NF-kB P65 binding activity and NFĸB p65 (pSer536) levels. These effects of DPGZ were accompanied by a significant reduction in NLRP3 levels and NLRP3 gene expression and a significant decrease in caspase-1 activity, which were also confirmed by histopathological examinations. We conclude that DPGZ antioxidant and anti-inflammatory activity may occur through regulation of AMPK/NFĸB pathway and inhibition of NLRP3 activation. These results suggest that DPGZ represents a promising intervention for the treatment of ALI, particularly in patients with type 2 diabetes.

Dual-Mode Gradient HPLC and TLC Densitometry Methods for the Simultaneous Determination of Paracetamol and Methionine in the Presence of Paracetamol Impurities.

BACKGROUND: Paracetamol (PC) is one of the most widely used analgesic and antipyretic drugs and has recently been integrated into the supportive therapy of COVID-19. Pharmaceuticals containing methionine (MT) with PC may contribute to avoid hepatotoxicity and eventual PC overdose-dependent death. OBJECTIVE: The current work purposes to develop and validate two chromatographic methods for the simultaneous determination of MT and PC in the presence of two PC impurities (4-nitrophenol [NP] and 4-aminophenol [AP]). METHOD: Two chromatographic methods were established and validated according to the International Conference on Harmonization guidelines. The first one was an RP-HPLC/UV method based on applying a "dual-mode" gradient elution. The separation was realized via varying both the composition of the ternary mobile phase (acetonitrile-methanol-water) and its flow rate. This strategy enabled a relatively rapid analysis with a satisfactory resolution, although the investigated compounds exhibit a significant difference in lipophilicity. The second one relied on TLC-densitometry, where the optimum separation was realized using a quaternary mobile phase system composed of butanol-dioxane-toluene-methanol (8:2.5:3.5:0.3, by volume). Both methods were monitored at 220 nm. RESULTS: The developed methods were proven to be robust, accurate, specific, and appropriate for the routine analysis of PC in its pure form or in pharmaceutical formulations with MT in quality control laboratories. CONCLUSIONS: The corresponding methods are suitable to determine MT and PC in the presence of PC impurities. HIGHLIGHTS: The study achieves the analysis of MT and PC in the presence of PC impurities via the application of HPLC and TLC-densitometry methods.

Simultaneous Determination of Paracetamol, Propyphenazone and Caffeine in Presence of Paracetamol Impurities Using Dual-Mode Gradient HPLC and TLC Densitometry Methods.

Two chromatographic methods were validated for the determination of the widely prescribed analgesic and antipyretic drug combination of paracetamol (PC) (recently integrated into the supportive treatment of COVID-19), propyphenazone (PZ) and caffeine (CF) in the presence of two PC impurities, namely 4-aminophenol and 4-nitrophenol. A "dual-mode" gradient high-performance liquid chromatography method was developed, where the separation was achieved via "dual-mode" gradient by changing both the ternary mobile phase composition (acetonitrile: methanol: water) and the flow rate. This enables a good resolution within a relatively shorter analysis time. The analysis was realized using Zorbax Eclipse XDB column C18, 5 µm (250 × 4.6 mm) and the UV detector was set at 220 nm. The other method is a thin-layer chromatography densitometry method, where the separation was achieved using a mobile phase composed of chloroform: toluene: ethyl acetate: methanol: acetic acid (6: 6: 1: 2: 0.1, by volume). Densitometric detection was performed at 220 nm on silica gel 60 F254 plates. The developed methods were fully validated as per the ICH guidelines and proved to be accurate, robust, specific and suitable for application as purity indicating methods for routine analysis of PC in pure form or in pharmaceuticals with PZ and CF in quality control laboratories.

Calixarene based portable sensor for the direct assay of indiscriminate ephedrine content of weight loss herbal preparations.

A novel potentiometric sensor was developed and optimized for the quantitative analysis of ephedrine in non-prescribed herbal supplements used as adjunctive therapy for weight loss. An initial optimization study aimed to reach the optimum membrane composition, sensor assembly, and experimental conditions. The study evaluated the effect of several factors on the sensor performance including different ion-exchangers, plasticizers, ionophores, membrane thicknesses, soaking solution concentrations, soaking time intervals, and pH. The optimized polyvinyl chloride membrane included tungstophosphoric acid hydrate as a cation exchanger, tricresyl phosphate as a plasticizer, and calix[8]arene as an ionophore to enhance the sensitivity and selectivity of the developed sensor. The polyvinyl chloride membrane was drop-casted over a polyaniline modified glassy carbon electrode surface to form a solid-state sensor. The proposed membrane succeeded to quantify ephedrine over a linear range of 6 × 10-6 to 1 × 10-2 M with a LOD of 3.60 × 10-6 M, acceptable selectivity, and fast response time. The IUPAC characterization of sensor response and International Conference on Harmonization validation parameters were calculated. The method successfully determined ephedrine concentration in spiked herbal mixtures and determined labeled and undeclared ephedrine content of weight loss herbal preparations.

Experimentally designed chromatographic method for the simultaneous analysis of dimenhydrinate, cinnarizine and their toxic impurities.

Recently, experimental design has beaten the traditional optimization approach (one variable at a time) by providing better quality for chromatographic separation using minimal effort and resources. Benzophenone (BZP) and [1-(diphenylmethyl)piperazine] (DPP) were reported to be the most toxic impurities for dimenhydrinate (DMH) and cinnarizine (CIN), respectively. Additionally, there is no reported HPLC method for the simultaneous determination of DMH, CIN and their toxic impurities. A custom experimental design was adopted to estimate the optimum conditions that achieved the most acceptable resolution with adequate peak symmetry within the shortest run time. Desirability function was used to define the optimum chromatographic conditions and the optimum separation was achieved using XBridge® HPLC RP-C18 (4.6 × 250 mm, 5 µm), acetonitrile: 0.1% sodium lauryl sulphate (SLS) in water (90 : 10, v/v) as a mobile phase at flow rate 2 mL min-1 and UV detection at 215 nm. Method validation was carried out according to ICH guidelines and linearity was achieved in the ranges of 2-25, 1-25, 1-12.5, and 1-12.5 µg mL-1 for DMH, CIN, BZP and DPP, respectively. By application of the proposed method to the market dosage form, no interference from excipients was observed. Moreover, the greenness of the method was evaluated using the National Environmental Method Index (NEMI), Analytical Eco-Scale and Green Analytical Procedure Index (GAPI) metrics and the results revealed the green environmental impact of the developed method.

Solid-state potentiometric sensor for the rapid assay of the biologically active biogenic amine (tyramine) as a marker of food spoilage.

Tyramine (TYR) is a vasoactive biogenic amine found in food products due to improper storage and poor hygiene. High TYR intake results in a wide range of life-threatening physiological reactions. The work optimizes a solid-state potentiometric sensor in the absence of a reported potentiometric method for rapid and direct TYR assay. The optimization study included thirteen membrane cocktails of different compositions. The optimized sensor proved a near-Nernstian slope of 57.30 mV/decade, a quantification limit of 10.6 ppm, and a detection limit of 7.9 ppm. Validation results confirmed the sensor ability for the direct assay of TYR in blue cheese, aged cheese, Egyptian pickled cottage cheese, and pickled herring. A comparison with the reported chromatographic method expresses the merits and potentials of the developed sensor for the rapid testing of food edibility, quality, and safety based on its TYR content. Chemical compounds studied in this article: Tyramine (PubChem CID: 5610); Tyramine hydrochloride (PubChem CID: 66449); Poly(vinyl chloride) (PubChem SID: 24864273); Tricresyl phosphate (PubChem CID: 6529); sodium phosphotungstate tribasic hydrate (PubChem SID: 329753864).

Chemotherapeutic and chemopreventive potentials of ρ-coumaric acid - Squid chitosan nanogel loaded with Syzygium aromaticum essential oil.

One of the most important trends in chemotherapy is the development of green chemotropic drugs with maximal activity and minimal side effects. The nanoencapsulation of phytochemical oils with natural polymers has been documented as a promising approach to producing nanodrugs with sustainable bioactivity and prolonged stability. In this context, Syzygium aromaticum essential oil (SAEO) and ultrasound-assisted deacetylated chitosan (UCS3) were successfully extracted from clove buds and squid pens, respectively. Grafting of UCS3 by ρ-coumaric acid (ρCA) has been performed to fabricate the ρCACS nanogel which was used for nanoencapsulation of SAEO to yield SAEO-loaded nanogel (ρCACS@SAEO). The findings of spectral, thermal, and morphological analyses have confirmed the success of the formation of new materials and SAEO encapsulation, as well. Based on the findings of the in vitro antimicrobial, antioxidant, and anticancer studies, the nanoencapsulation of SAEO by ρCACS has significantly boosted its chemotherapeutic effects, compared to unencapsulated oil. Therefore, ρCACS@SAEO nanogel could be considered as a multifunctional chemotherapeutic/chemopreventive agent for prevention or therapy of pathologies induced by oxidative stress, microbial infection, and breast and skin cancer.

Evaluation of Synthetic 2,4-Disubstituted-benzo[g]quinoxaline Derivatives as Potential Anticancer Agents.

A new series of 2,4-disubstituted benzo[g]quinoxaline molecules have been synthesized, using naphthalene-2,3-diamine and 1,4-dibromonaphthalene-2,3-diamine as the key starting materials. The structures of the new compounds were confirmed by spectral data along with elemental microanalyses. The cytotoxic activity of all synthesized benzo[g]quinoxaline derivatives was assessed in vitro against the breast MCF-7 cancer cell line. The tested molecules revealed good cytotoxicity toward the breast MCF-7 cancer cell line, especially compound 3. The results of topoisomerase IIß inhibition assay revealed that compound 3 exhibits potent inhibitory activity in submicromolar concentration. Additionally, compound 3 was found to cause pre-G1 apoptosis, and slightly increase the cell population at G1 and S phases of the cell cycle profile in MCF-7 cells. Finally, compound 3 induces apoptosis via Bax activation and downregulation of Bcl2, as revealed by ELISA assay.