F1000Research F1000Research 2046-1402 F1000 Research Limited London, UK 10.12688/f1000research.172260.1 Research Article Articles Evaluation of the effectiveness of hesperidin and its derivatives in inhibiting human cancer MCF-7 cells and leshmania: an in vitro study

[version 1; peer review: awaiting peer review]

 Al-Halbosiy Mohammad Formal Analysis Methodology Project Administration Supervision Writing – Review & Editing https://orcid.org/0000-0003-1373-4188 a 1 Al Shmgani Hanady Writing – Original Draft Preparation https://orcid.org/0000-0002-9108-4603 b 2 Al-Jumaily Rakad Data Curation Investigation 3 Abdulhameed Baraa Methodology 1 Jaed Lamees Resources Validation Writing – Original Draft Preparation 2 Al-Nahrain University/ Biotechnology research center, Baghdad, Iraq Biology Department, College of Education for Pure Sciences/Ibn al-Haitham, University of Baghdad, Baghdad ,Iraq, Baghdad, Iraq Department of Biology, College of Science, University of Baghdad, Baghdad, Iraq, Baghdad, Iraq mohammad.farhan@nahrainuniv.edu.iq Hanadi.s.as@ihcoedu.uobaghdad.edu.iq

No competing interests were disclosed.

 16 1 2026 2026 15 69 5 1 2026 Copyright: © 2026 Al-Halbosiy M et al. 2026 This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Background

Natural resources and their pharmaceutical products play an important role in disease management because of their potential for new drug discovery and historical traditional use in healthcare. This study aimed to investigate the effects of hesperidin and its derivatives on MCF-7 human breast cancer cells and leishmaniosis.

 Methods

The colorimetric cell viability MTT assay was used to measure the efficacy of these compounds in inhibiting MCF-7 cell growth and the promastigote forms of Leishmania tropica. Cells treated with different concentrations (3.125, 6.25, 12.5, 25, 50, 100, and 200 μg/ml) of hesperidin and their derivative, derv.1(5-amino-1,3,4-thiadiazole-2-thiol), and derv. 2 (5-(methylthio)-1,3,4-thiadiazol-2-thiol) after 24 h exposure.

 Results

The Results showed that hesperidin was the most effective in inhibiting MCF-7 cells, with a 58.3% inhibition rate at a concentration of 100 μg/ml, whereas the derivative (derv.2) exhibited the highest antileishmanial activity, with a 50.2% inhibition rate at the same concentration. The anticancer and antileishmanial effects were concentration-dependent, with inhibition rates decreasing at lower concentrations.

 Conclusions

These findings indicate the potential role of hesperidin and its derivatives as anti-cancer and antileishmanial agents, suggesting new ways for further research into their medical applications.

 hesperidin anti-cancer antileishmanial MCF-7 human breast cancer The author(s) declared that no grants were involved in supporting this work. Introduction

Natural resources and their pharmaceutical products play an important role in the management of diseases with almost no adverse effects. Many studies have revealed that natural substances or their main components can enhance health in different ways, such as decreasing inflammation and oxidative stress, managing diseases, including autoimmune disease and cancer, and regulating the pathways of cell signaling. 1 The utilization of agriculture and/or its waste products for producing bioactive products is gaining traction because of its cost-effectiveness and potential to address waste disposal issues. This approach offers a sustainable alternative, as highlighted by research focusing on environmental implications. 2 Hesperidin (HSP) is a bioflavonoid found mainly in citrus fruits, particularly in peels. It functions as a protective agent and is produced by secondary plant metabolism. It exhibits various biological activities, including antioxidant, antibacterial, anti-inflammatory, and antitumor effects. Beyond citrus, HSP is also present in plant genera, such as Lamiaceae, Fabaceae, and Betulaceae. Its classification as a glycoside flavone underscores its role in plant defense and potential health benefits. 3, 4

Cancer is characterized by uncontrolled division of cells, leading to the formation of tumors that can invade other tissues and metastasize at the speed at which cancer develops and becomes dangerous varies, with some cancers progressing rapidly while others take years. Treatments often target uncontrolled growth of cells. Breast cancer, the most common cancer in women, has a significant impact on mortality and morbidity. 5, 6 Many studies have confirmed the role of natural products in preventing the development of breast cancer by promoting cancer cell apoptosis, inhibiting proliferation and metastasis of cancer cells, and avoiding signal transduction. 7, 8 However, many biomolecules found in the human diet, including flavonoids, can act as chemopreventers and reduce cancer cell growth. Many mechanisms have been suggested to explain the effect of these biomolecules, such as apoptosis and cell-cycle arrest, in addition to their antioxidant qualities. 9

Leishmaniasis is a parasitic disease caused by protozoan parasites of the Leishmania genus, transmitted through the bite of infected female phlebotomine sand flies. It is a neglected tropical disease (NTD) with a wide spectrum of clinical manifestations, ranging from self-healing cutaneous lesions to life-threatening visceral forms. 10 The disease is endemic in tropical and subtropical regions and remains a significant public health challenge, particularly in resource-limited settings requiring integrated control strategies to reduce its burden.

Hesperidin, as well as many natural flavonoids, show low water solubility and few lipophilic molecules, hindering their ability to cross the cell membrane. To overcome these limitations, in our previous study, novel HSP derivatives were developed and synthesized that exhibited promising anticytotoxic and antioxidant properties. 11 In this study, we investigated the effects of HSP and its derivatives on MCF-7 human breast cancer cells, and their potential efficacy against leishmaniasis.

To overcome these limitations, new HSP derivatives have been developed and synthesized. Two novel HSP derivatives were successfully synthesized and described in our previous study, which exhibited promising anticytotoxic and antioxidant activities. 11 This study investigated the effects of hesperidin and its derivatives on MCF-7 human breast cancer cells and on leishmaniosis.

 Materials and methods Chemicals

Hesperidin (Catalogue no. H5006; ≥80%), and 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) (Catalogue no. 475989) were purchased from Sigma Aldrich (St. Louis, MO, USA). Roswell Park Memorial Institute (RPMI) 1640 medium, fetal bovine serum (FBS), and antibiotic–antimycotic solution were procured from Gibco (Gaithersburg, MD, USA).

 Preparation and synthesis of HSP derivatives

The two derivatives used in this study were prepared and synthesized as described in our previous study. 11 The purity of the recrystallized compounds was assessed via thin-layer chromatography (TLC), and their structure was verified through physical properties and spectral analyses. The open capillary tube method was used to determine the melting point of the samples. IR spectra were obtained using Fourier-Transform Infrared (FTIR) spectroscopy and H-NMR spectra were acquired on a Bruker AVN 300 MHz spectrometer. Electron impact mass spectrometry (EIMS) analysis was performed using a Shimadzu-GC-MS-QP2010 Ultra System.

A solution of the first derivative, [drev.1] (5-amino-1,3,4-thiadiazole-2-thiol), was added dropwise to the HSP solution (0.133 g, 1 mmol) in glacial acetic acid (10 ml). The reaction mixture was refluxed for 18 h (monitored by TLC, CHCL 3, MeOH 1:1). The resulting precipitate was collected, washed with water, and recrystallized from aqueous DMF to yield a white amorphous solid (78%, mp 219-221°C). The second derivative was prepared as above with some medication (HSP 5-(methylthio)-1,3,4-thiadiazol-2-thiol [drev.2]; the modifications included thioether linkage at the 5-position of thiadiazole ring, shorter reaction time to 12 h, ethanol recrystallization solvent, and melting point 205-207°C.

 Anticancer activity

The percentage of breast cancer growth inhibition was determined using the colorimetric MMT assay. The MCF-7 cell line (Michigan Cancer Foundation 7) is a widely used human breast cancer model, derived from a 69-year-old woman’s metastatic adenocarcinoma, we obtained it from the biotechnology research center/Al-Nahrain University, Baghdad, Iraq. Briefly, MCF-7 cells were seeded at a density of 1×10 6 cells/ml in a 96-well plate. The test compound (hesperidin) was derived. 1 and derive. 2) were dissolved in an aqueous solution at concentrations of 3.15, 6.25, 12.5, 25, 50, and 100 μg/ml. Subsequently, 100 μL of each concentration was added to the respective well, followed by incubation at 37°C for 24 h. After incubation, 10 μL MTT solution (5 mg/ml) was added to each well and incubated for an additional 4h. To solubilize the formazan crystals, 50 μL of dimethyl sulfoxide (DMSO) was added to each well, followed by a 10-minute incubation. Untreated cell culture medium served as a negative control. All experiments were conducted in triplicates. The absorbance was measured at 620 nm using an ELISA reader, and the mean absorbance values for each replicate group were calculated. Cell viability (expressed as a percentage) and the inhibition ratio were determined using standard formulae, as follows 12 : IG % = ( Ac − As / Ac ) × 100

Ac = optical density of control, As = optical density of samples.

 Antileishmanial activity assessment

The antileishmanial activity of hesperidin and its derivatives (derive.1 and derive.2) was evaluated against the promastigote form of Leishmania tropica. 13 The L. tropica strain was obtained from and maintained at the Biotechnology Research Center, Al-Nahrain University. Promastigotes were cultured in RPMI-1640 medium supplemented with L-glutamine (20 mM), 10% fetal bovine serum, 100 U/ml penicillin, and 50 μg/ml streptomycin, all treated samples then incubated at 22-26°C.

Cell viability was assessed using the colorimetric MTT assay, as described in anticancer section above. L. tropica promastigotes (1 × 10 6 cells/mL, 100 μL/well) were seeded into a 96-well tissue culture plate. The test compounds (hesperidin and Derv. 1, and Derv. 2) were dissolved in aqueous solutions at concentrations of 3.125, 6.25, 12.5, 25, 50, and 100 μg/mL, respectively. Subsequently. The following equation was used to determine cell growth inhibition: IG % = ( Ac − As / Ac ) × 100

Where AC = optical density of control, As = optical density of samples.

 Statistical analysis

The experimental data were analyzed using SPSS (SPSS 23.0, SPSS Ins. III., USA). Significant differences were assessed using One-Way Analysis of variance (ANOVA), and P values ≤ 0.05 were considered significant.

 Results and Discussion

MTT assay was used to measure the viability of MCF-7 breast cancer cells. Cells were treated with different concentrations (3.125, 6.25, 12.5, 25, 50, and 100 μg/ml) of hesperidin and its derivatives (derv.1 and 2) for 24 h. The Results of hesperidin exposure showed the best percentage of inhibition at 48.9% and 57.3% at the highest concentrations (50 and 100 μg/ml), respectively, compared to derv.1 (39.3% and 37.5%) and derv.2 (36.8% and 35%), respectively, as shown in Table 1. Also a considerable variance reveled between the 25 and 12.5 μg/ml with the lowest concentrations (6.25 and 3.125 μg/ml) among the different of hesperidin and their derv.1 and 2 ( Table 1).

Percentage of inhibition of MCF-7 cell line by using the hesperidin and their derivative (derv.1 and 2) during 24 hours of exposure.
Conc. μg/ml Hesperidin Hesperidin derv.1 Hesperidin derv.2
Mean ± SD GI% Mean ± SD GI% Mean ± SD GI%
100 0.196 ± 0.002 57.39 0.139 ± 0.057 39.3 0.144 ± 0.038 36.8
50 0.234 ± 0.018 48.9 0.143 ± 0.003 37.5 0.148 ± 0.007 35
25 0.262 ± 0.058 43 0.149 ± 0.006 34.9 0.155 ± 0.011 32
12.5 0.293 ± 0.039 36.3 0.178 ± 0.013 22.1 0.185 ± 0.019 18.8
6.25 0.376 ± 0.016 18.3 0.190 ± 0.004 17 0.197 ± 0.003 13.5
3.125 0.419 ± 0.140 8.9 0.212 ± 0.044 7.4 0.218 ± 0.024 4.4
Control 0.460 ± 0.002 - 0.229 ± 0.013 - 0.228 ± 0.019 -

-No inhibition.

The results in Table 2 indicate the antileishmanial activity of hesperidin and its derivative (derv.1 and 2) against promastigote forms of Leishmania tropica. Cells were treated with different concentrations (3.125, 6.25, 12.5, 25, 50, and 100 μg/ml) of hesperidin and derv.1 and 2 during 24 h of exposure. The Results of derv.2 exposure showed the highest percentage of inhibition of 35.7% and 50.2% at the highest concentrations (50 and 100 μg/ml), respectively, in comparison to hesperidin (30.1% and 24.7%) and derv.1 (33.3% and 25.2%), respectively, as shown in Table 2. In addition, considerable variance was observed at 25 μg/ml concentration between derv.2 (32.6%) and hesperidin (16%) and derv.1 (17.6%). However, no differences were observed at the lowest concentrations (6.25 and 3.125 μg/ml) among the hesperidin and their derv.1 and 2 ( Table 2).

Percentage of inhibition of promastigote forms of Leishmania tropica by using the hesperidin and their derivative (derv.1 and 2) during 24 hours of exposure.
Conc. μg/ml Hesperidin Hesperidin derv.1 Hesperidin derv.2
Mean ± SD GI% Mean ± SD GI% Mean ± SD GI%
100 0.304 ± 0.007 30.3 0.132 ± 0.002 33.3 0.230 ± 0.019 50.2
50 0.328 ± 0.049 24.7 0.148 ± 0.006 25.2 0.297 ± 0.005 35.6
25 0.366 ± 0.019 16 0.163 ± 0.007 17.6 0.311 ± 0.011 32.6
12.5 0.395 ± 0.009 9.4 0.180 ± 0.001 9 0.378 ± 0.005 18.2
6.25 0.412 ± 0.011 5.5 0.189 ± 0.005 4.5 0.421 ± 0.029 8.8
3.125 0.485 ± 0.026 0 0.201 ± 0.004 0 0.478 ± 0.001 0
Control 0.436 ± 0.022 - 0.198 ± 0.004 - 0.462 ± 0.002 -

-No inhibition.

The investigation of natural products and their semi-synthetic derivatives remains the cornerstone of pharmaceutical discovery, offering a rich reservoir of chemical scaffolds with diverse biological activities. This study contributes to this field by evaluating the anticancer and antileishmanial potential of the flavonoid hesperidin and two of its synthesized thiadiazole derivatives. These findings provide compelling preliminary evidence that the structural modification of a natural compound can selectively enhance its bioactivity against different pathogenic targets.

The most potent anticancer activity against MCF-7 human breast cancer cells was exhibited by the parent compound, hesperidin, which achieved 58.3% inhibition at 100 μg/ml with concentration-dependent inhibition of MCF-7 cell growth by hesperidin. This observation aligns with a substantial body of literature documenting the chemopreventive and antitumor properties of flavonoids by inducing apoptosis and inhibiting proliferation in cancer cells through mechanisms such as cell cycle arrest, often at the G1/S or G2/M checkpoints, and exhibiting potent antioxidant and anti-inflammatory effects that indirectly suppress tumor promotion and progression. 14 The superior efficacy of HSP over its derivatives (derv.1 and 2) in inhibiting MCF-7 cells suggests that the pure structure may be more conducive to interacting with cancer cell pathways, possibly with its glycoside flavone backbone. 15 However, the fact that the thiadiazole derivatives (derv.1 and derv.2) showed lower efficacy against MCF-7 cells than the parent hesperidin is a critical finding. This suggests that the specific structural motifs of hesperidin, particularly its glycosylated rutinose moiety and characteristic flavanone skeleton, are crucial for its interaction with molecular targets in breast cancer cells. The introduction of the 1,3,4-thiadiazole ring, while potentially beneficial for other biological activities, may have altered the pharmacokinetics of the molecule (e.g., reducing cellular uptake) or its affinity for key oncogenic targets such as protein kinases or estrogen receptors to which MCF-7 cells are responsive. Currently, anticancer research on natural compounds has focused on the induction of apoptosis in cancer cells. 16 Hesperidin induces apoptosis in cancer cells through the intrinsic mitochondrial pathway, characterized by the upregulation of pro-apoptotic proteins (e.g., Bax) and downregulation of anti-apoptotic proteins (e.g., Bcl-2). 17 Siddiqui et al. 18 indicated that hesperidin has a promising impact on the treatment of cancer due to its high efficacy and fewer side effects than non-natural drugs. The concentration-dependent effect observed in both assays was consistent with the pathway principles of natural compounds, where higher doses typically produce higher therapeutic results. 19 However, the decline in efficacy at lower concentrations highlights the need for further optimization to enhance potency, while minimizing potential side effects. Moreover, other studies in which minor modifications significantly altered compound efficacy. Moreover, studies have suggested some mechanisms of HSP and its derivatives in anticancer and antileishmanial activities, such as induction of apoptosis through mitochondrial dysfunction or modulation of signaling pathways via PI3K/AKT and MAPK. 20, 21

Enhanced antileishmanial activity of derivative derv.2

In contrast to the anticancer results, the derivative derv.2 (5-(methylthio)-1,3,4-thiadiazol-2-thiol) demonstrated antileishmanial activity against the promastigote forms of Leishmania tropica. This significant observation underscores the principle of selective bioactivity through rational chemical modifications. The 1,3,4-thiadiazole nucleus is a privileged structure in medicinal chemistry that is renowned for its broad-spectrum antimicrobial, antifungal, and antiparasitic properties. The results showed that derv.2 was the most effective, as it achieved a 50.2% inhibition rate at 100 μg/m, outperforming both HSP and derv.1. This suggests that the thioether linkage at the 5-position of the thiadiazole ring in derv.2 may enhance its activity against Leishmania by improving membrane permeability or targeting specific enzymes. 22 Increasing lipophilicity is another suggested mechanism: the methylthio (-SCH3) group in derv.2 likely increases the lipophilicity of the compounds compared to polar glycosylated hesperidin and amino-substituted derv.1. This enhanced lipophilicity facilitates passive diffusion across the unique and complex cell membrane of the leishmania parasite, which is rich in glycosylphosphatidylinositols and other lipids. 23 The fact that derv.2 was more effective against the parasite than the parent compound suggests the potential for selective toxicity. The unique biochemical pathways in leishmania (e.g., the trypanothione system) present targets that are absent in human cells, which could be more susceptible to the novel thiadiazole derivative. 24

The consistent observation of concentration-dependent anticancer and antileishmanial activities strengthens the validity of the findings. This indicated that the effects were specific and not due to general cytotoxicity at high concentrations. The divergent structure-activity relationship revealed that hesperidin is optimal for anticancer activity, and its thiadiazole derivative (derv.2) is superior for antileishmanial activity is of paramount importance. This suggests that a single natural product can serve as a platform for generating multiple lead compounds optimized for distinct therapeutic indications.

In conclusion, this study highlights the dual potential of HSP and derv. (1) and (2) as anticancer and antileishmanial agents, respectively. While HSP shows promise against MCF-7 cells, its derivative derv.2 emerges as a more effective antileishmanial candidate. These findings pave the way for further research on structural optimization, mechanistic studies, and preclinical trials to develop these compounds as viable therapeutic options.

 Data availability

Zenodo. Evaluation of the effectiveness of hesperidin and its derivatives in inhibiting human cancer MCF-7 cells and leshmania: an in vitro study. https://doi.org/10.5281/zenodo.17793855. 25

This project contains the following underlying data

Table 1: Raw data Excel sheet. The values behind the means, standard deviations and other measures reported in MCF-7 cell line percentage inhibition by hesperidin and their derivatives

Table 2: Raw data Excel sheet. The values behind the means, standard deviations and other measures reported in Leishmania tropica cell line percentage inhibition by hesperidin and their derivatives.

Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0).

 References Anywar GU Kakudidi E Oryem-Origa H : Cytotoxicity of Medicinal Plant Species Used by Traditional Healers in Treating People Suffering From HIV/AIDS in Uganda. Front. Toxicology. 2022;4:832780. 35586188 10.3389/ftox.2022.832780 PMC9108544 Guo J Zhang Y Fang J : Reduction and Reuse of Forestry and Agricultural Bio-Waste through Innovative Green Utilization Approaches: A Review. Forests. 2024;15(8):1372. 10.3390/f15081372 Ogunro OB : An updated and comprehensive review of the health benefits and pharmacological activities of hesperidin. Biochem. Biophys. Res. Commun. 2025 Jul 30;772:151974. Epub 2025 May 11. 40414011 10.1016/j.bbrc.2025.151974 Al-Kubaisi ZA Al-Shmgani HS Salman MJ : Evaluation of in vivo and in vitro protective effects of quercetin on lipopolysaccharide-induced inflammation and cytotoxicology. Research Journal of Pharmacy and Technology. 2020;13(8):3897–3903. 10.5958/0974-360X.2020.00690.3 Cooper GM : The Cell: A Molecular Approach. Sunderland (MA): Sinauer Associates; 2nd edition 2000. The Development and Causes of Cancer. Reference Source Sahar BR Al-Jumaily RMK : The Impact of Global DNA Methylation and Hypoxia-Inducible Factor 1 Alpha Levels in the Progression of Breast Cancer. OM&P. 2024;11(2):28–39. Al-Rikabi RH Al-Shmgani HS : Evaluation of hesperidin protective effect on lipopolysaccharide-induced inflammation and lipid peroxidation in BALB/C male mice. Res. J. Pharm. Tech. 2018;11:5513–5516. Hussain T Bajpai S Saeed M : Potentiating effect of ethnomedicinal plants against proliferation on different cancer cell lines. Curr. Drug Metab. 2018;19:584–595. 29512451 10.2174/1389200219666180305144841 Di Dalmazi G Giuliani C Bucci I : Promising Role of Alkaloids in the Prevention and Treatment of Thyroid Cancer and Autoimmune Thyroid Disease: A Comprehensive Review of the Current Evidence. Int. J. Mol. Sci. 2024;25:5395. Vries HJC de Schallig HD : Cutaneous Leishmaniasis: A 2022 Updated Narrative Review into Diagnosis and Management Developments. Am. J. Clin. Dermatol. 2022;23:823–840. 36103050 10.1007/s40257-022-00726-8 PMC9472198 Al-Shmgani H Shakir R Wahed A : Design, Synthesis, Docking, Antitumor Screening, And Absorption, Distribution, Metabolism, And Excretion Prediction Of New Hesperdin Derivative. Asian J. Pharm. Clin. Res. 2020;13:24–31. Makia R Al-Sammarrae K Al-Halbosiy M : In Vitro Cytotoxic Activity of Total Flavonoid from Equisetum Arvense Extract. Rep. Biochem. Mol. Biol. 2022 Oct;11(3):487–492. 36718311 10.52547/rbmb.11.3.487 PMC9883023 Das A Das MC Das N : Evaluation of the antileishmanial potency, toxicity and phytochemical constituents of methanol bark extract of Sterculia villosa. Pharm. Biol. 2017 Dec;55(1):998–1009. 28173714 10.1080/13880209.2017.1285946 PMC6130613 Rahmani AH Babiker AY Anwar S : Hesperidin, a Bioflavonoid in Cancer Therapy: A Review for a Mechanism of Action through the Modulation of Cell Signaling Pathways. Molecules. 2023 Jun 30;28(13):5152. 37446814 10.3390/molecules28135152 PMC10343639 Pyrzynska K : Hesperidin: A Review on Extraction Methods, Stability and Biological Activities. Nutrients. 2022;14(12):2387. 35745117 10.3390/nu14122387 PMC9227685 Roohbakhsh A Parhiz H Soltani F : Neuropharmacological properties and pharmacokinetics of the citrus flavonoids hesperidin and hesperetin – a mini-review. Life Sci. 2014;113:1–6. 25109791 10.1016/j.lfs.2014.07.029 Aggarwal V Tuli HS Thakral F : Molecular mechanisms of action of hesperidin in cancer: Recent trends and advancements. Exp. Biol. Med. (Maywood). 2020 Mar;245(5):486–497. 32050794 10.1177/1535370220903671 PMC7082885 Siddiqui IA Sanna V : Impact of nanotechnology on the delivery of natural products for cancer prevention and therapy. Mol. Nutr. Food Res. 2016;60:1330–1341. 26935239 10.1002/mnfr.201600035 Onufrak NJ Forrest A Gonzalez D : Pharmacokinetic and Pharmacodynamic Principles of Anti-infective Dosing. Clin. Ther. 2016 Sep;38(9):1930–1947. 27449411 10.1016/j.clinthera.2016.06.015 PMC5039113 Jeong SA Yang C Song J : Hesperidin Suppresses the Proliferation of Prostate Cancer Cells by Inducing Oxidative Stress and Disrupting Ca 2+ Homeostasis. Antioxidants. 2022;11:1633. 36139707 10.3390/antiox11091633 PMC9495577 Lee KH Yeh MH Kao ST : The inhibitory effect of hesperidin on tumor cell invasiveness occurs via suppression of activator protein 1 and nuclear factor-kappaB in human hepatocellular carcinoma cells. Toxicol. Lett. 2010;194:42–49. 20138977 10.1016/j.toxlet.2010.01.021 Nandikolla A Srinivasarao S Karan Kumar B : Synthesis, study of antileishmanial and antitrypanosomal activity of imidazo pyridine fused triazole analogues. RSC Adv. 2020 Oct 19;10(63):38328–38343. 35517538 10.1039/d0ra07881f PMC9057266 Liu X Testa B Fahr A : Lipophilicity and its relationship with passive drug permeation. Pharm. Res. 2011 May;28(5):962–977. 21052797 10.1007/s11095-010-0303-7 Brígido HPC Santos LGA dos Barros RC de : The Role of Oxidative Stress in the Pathogenesis and Treatment of Leishmaniasis: Impact on Drug Toxicity and Therapeutic Potential of Natural Products. Toxics. 2025;13(3):190. 40137517 10.3390/toxics13030190 PMC11945835 Al-Halbosiy M Al-Shmgani H Al-Jumaily R : Evaluation of the effectiveness of hesperidin and its derivatives in inhibiting human cancer MCF-7 cells and leshmania: an in vitro study. Zenodo. 2025. 10.5281/zenodo.17793855