Newly Synthesized Imidazoline Compound Regulates Apoptosis and Toll-Like Receptor Signaling in Lung Adenocarcinoma and Normal Epithelial Cells

Öz

Imidazoline is a heterocyclic compound with biological activity in regulating apoptosis, inflammation, and immune signaling. Lung cancer is a leading cause of cancer-related death, emphasizing the need for new therapies. This study examined the effects of imidazoline on apoptosis and Toll-like receptor (TLR) expression in A549 lung adenocarcinoma cells and Beas-2B normal bronchial epithelial cells. The chemical structure of the synthesized imidazoline was confirmed by FT-IR spectroscopy. Cells were treated with imidazoline, and expression of apoptotic markers (BAX, BCL-2) and TLR1-10 genes was analyzed. In A549 cells, imidazoline increased BAX and decreased BCL-2, indicating activation of intrinsic apoptosis and cell cycle arrest. Beas-2B cells showed the opposite pattern, suggesting selective toxicity toward cancer cells. Imidazoline also modulated several TLRs in both cell lines, highlighting potential immunoregulatory effects. These findings demonstrate that imidazoline promotes apoptosis and alters immune-related signaling in lung cancer cells without harming normal epithelial cells. This study is the first to report imidazoline’s impact on TLR expression in lung cancer, supporting its potential as a dual anticancer agent targeting both cell death and immune pathways.

Anahtar Kelimeler

• İmidazoline
• Lung cancer
• Toll-like receptors
• Apoptosis

Destekleyen Kurum

No

Etik Beyan

Ethical approval is not required.

Teşekkür

The authors would like to thank NF Chemical for their support in the synthesis and characterization of the imidazoline compound used in this study.

Yeni Sentezlenen İmidazolin Bileşiği, Akciğer Adenokarsinomunda ve Normal Epitel Hücrelerinde Apoptosisi ve Toll-Like Reseptör Sinyallemesini Düzenler

Öz

İmidazolin, apoptosis, inflamasyon ve bağışıklık sinyal yollarını düzenleyen biyolojik aktiviteye sahip heterosiklik bir bileşiktir. Akciğer kanseri, kanser kaynaklı ölümler arasında önde gelen nedenlerden biridir ve yeni tedavi stratejilerine ihtiyaç olduğunu göstermektedir. Bu çalışmada, imidazolinin A549 akciğer adenokarsinomu hücrelerinde ve Beas-2B normal bronş epitelyum hücrelerinde apoptosis ve Toll-like reseptör (TLR) ekspresyonu üzerindeki etkileri incelenmiştir. Sentezlenen imidazolin bileşiğinin kimyasal yapısı FT-IR spektroskopisi ile doğrulanmıştır. Hücreler imidazoline ile muamele edilmiş ve apoptotik belirteçler (BAX, BCL-2) ile TLR1-10 genlerinin ekspresyonu analiz edilmiştir. A549 hücrelerinde imidazolin BAX seviyesini artırıp BCL-2 seviyesini düşürerek intrinsic apoptosis ve hücre döngüsü duraklamasını indüklemiştir. Beas-2B hücrelerinde ise ters bir desen gözlenmiş, bu da ilacın kanser hücrelerine seçici etkisini göstermiştir. Ayrıca, imidazolin her iki hücre hattında bazı TLR’leri modüle ederek potansiyel immün düzenleyici etkilerini ortaya koymuştur. Sonuçlar, imidazolinin kanser hücrelerinde apoptosis ve bağışıklıkla ilişkili sinyal yollarını etkileyerek normal hücrelere zarar vermeden çift yönlü antikanser aktivite gösterebileceğini ortaya koymaktadır. Bu çalışma, imidazolinin akciğer kanserinde TLR ekspresyonu üzerindeki etkilerini bildiren ilk çalışmadır ve bileşiğin hem hücre ölümü hem de immün yolları hedefleyen potansiyel bir antikanser ajan olduğunu desteklemektedir.

Anahtar Kelimeler

• İmidazolin
• Akciğer kanseri
• Toll benzeri reseptörler
• Apoptoz

Destekleyen Kurum

Yok

Etik Beyan

Etik onay gerektirmemektedir.

Teşekkür

Yazarlar, bu çalışmada kullanılan imidazoline bileşiğinin sentezi ve karakterizasyonundaki destekleri için NF Chemical'e teşekkür etmek isterler.

Kaynakça

1. Hanahan D. Hallmarks of cancer: new dimensions. Cancer Discov. 2022;12:31-46. https://doi.org/10.1158/2159-8290.CD-21-1059
2. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2021;71:209-49. https://doi.org/10.3322/caac.21660
3. Leiter A, Veluswamy RR, Wisnivesky JP. The global burden of lung cancer: current status and future trends. Nat Rev Clin Oncol. 2023;20:624-39. https://doi.org/10.1038/s41571-023-00798-3
4. Sağ S, Bayav İ, Tomatır AG, Dodurga Y. Palbociclib induces apoptosis and modulates toll-like receptor expression in lung adenocarcinoma and epithelial cells. Bratisl Med J. 2025;1-9. https://doi.org/10.1007/s44411-025-00319-x
5. Alduais Y, Zhang H, Fan F, Chen J, Chen B, et al. Non-small cell lung cancer (NSCLC): a review of risk factors, diagnosis, and treatment. Medicine. 2023;102(8):e32899. https://doi.org/10.1097/MD.0000000000032899
6. Lahiri A, Maji A, Potdar PD, et al. Lung cancer immunotherapy: progress, pitfalls, and promises. Mol Cancer. 2023;22:40. https://doi.org/10.1186/s12943-023-01740-y
7. Yousuf M, Shamsi A, Queen A, Shahbaaz M, Khan P, Hussain A, et al. Targeting cyclin‐dependent kinase 6 by vanillin inhibits proliferation of breast and lung cancer cells: combined computational and biochemical studies. J Cell Biochem. 2021;122:897-910. https://doi.org/10.1002/jcb.29921
8. Krasavin M. Biologically active compounds based on the privileged 2-imidazoline scaffold: the world beyond adrenergic/imidazoline receptor modulators. Eur J Med Chem. 2015;97:525-37. https://doi.org/10.1016/j.ejmech.2014.11.028

9. Sharma P, LaRosa C, Antwi J, Govindarajan R, Werbovetz KA. Imidazoles as potential anticancer agents: an update on recent studies. Molecules. 2021;26:4213. https://doi.org/10.3390/molecules26144213
10. Goel R, Luxami V, Paul K. Imidazo[1,2-a]pyridines: promising drug candidate for antitumor therapy. Curr Top Med Chem. 2016;16:3590-3616. https://doi.org/10.2174/1568026616666160414122644
11. Kowalski S, Wityk P, Raczak-Gutknecht J, Olszewska A, Żmijewski M, Kocić I, et al. The imidazoline I2 receptor agonist 2-BFI enhances cytotoxic activity of hydroxychloroquine by modulating oxidative stress, energy-related metabolism and autophagic influx in human colorectal adenocarcinoma cell lines. Eur J Pharmacol. 2025;996:177590. https://doi.org/10.1016/j.ejphar.2025.177590
12. Arnoux A, Aubertin G, Da Silva S, Weiss M, Bousquet P, Monassier L, et al. Nischarin is not the functional I1 imidazoline receptor involved in blood pressure regulation. J Cardiovasc Pharmacol. 2022;79:229-34. https://doi.org/10.1097/FJC.0000000000001128
13. Amisten S, Duner P, Asplund O, Mohammed Al-Amily I, Groop L, Salehi A. Activation of imidazoline receptor I2, and improved pancreatic β-cell function in human islets. J Diabetes Complications. 2018;32:813-18. https://doi.org/10.1016/j.jdiacomp.2018.06.010
14. Dickerson MT, Dadi PK, Zaborska KE, et al. Gi/o protein-coupled receptor inhibition of beta-cell electrical excitability and insulin secretion depends on Na+/K+ ATPase activation. Nat Commun. 2022;13:6461. https://doi.org/10.1038/s41467-022-34166-z
15. Bayav I, Sağ S, Ergezgin H, Dodurga Y, Tomatır AG. Possible TLR-mediated immunostimulatory effect of Palbociclib in breast cancer. Mol Biol Rep. 2026;53:120.
16. Keshavarz A, Pourbagheri-Sigaroodi A, Zafari P, Bagheri N, Ghaffari SH, Bashash D. Toll-like receptors (TLRs) in cancer; with an extensive focus on TLR agonists and antagonists. IUBMB Life. 2021;73:10-25. https://doi.org/10.1002/iub.2412
17. Aluri J, Cooper MA, Schuettpelz LG. Toll-like receptor signaling in the establishment and function of the immune system. Cells. 2021;10:1374. https://doi.org/10.3390/cells10061374
18. Mokhtari Y, Pourbagheri-Sigaroodi A, Zafari P, Bagheri N, Ghaffari SH, Bashash D. Toll-like receptors (TLRs): an old family of immune receptors with a new face in cancer pathogenesis. J Cell Mol Med. 2021;25:639-51. https://doi.org/10.1111/jcmm.16214
19. Pahlavanneshan S, Sayadmanesh A, Ebrahimiyan H, Basiri M. Toll-like receptor-based strategies for cancer immunotherapy. J Immunol Res. 2021;2021:9912188. https://doi.org/10.1155/2021/9912188
20. Rolfo C, Giovannetti E, Martinez P, et al. Applications and clinical trial landscape using Toll-like receptor agonists to reduce the toll of cancer. npj Precis Oncol. 2023;7:26. https://doi.org/10.1038/s41698-023-00364-1
21. Tyagi R, Tyagi VK, Pandey SK. Imidazoline and its derivates: an overview. J Oleo Sci. 2007;56:211-22.
22. Li Y, Yan B, He S. Advances and challenges in the treatment of lung cancer. Biomed Pharmacother. 2023;169:115891.
23. Hendriks LEL, Remon J, Faivre-Finn C, et al. Non-small-cell lung cancer. Nat Rev Dis Primers. 2024;10:71. https://doi.org/10.1038/s41572-024-00551-9
24. Adams SJ, Stone E, Baldwin DR, Vliegenthart R, Lee P, Fintelmann FJ. Lung cancer screening. Lancet. 2023;401:390-408.
25. Ayna A, Sağ S, Bayav İ, Darendelioğlu E. Chrysin protects neuronal cells against carboplatin exposure-induced apoptosis and oxidative damage. J Cell Neurosci Oxid Stress. 2025;17:1237-44.
26. Sriplai N, Sombatmankhong K. Corrosion inhibition by imidazoline and imidazoline derivatives: a review. Corros Rev. 2023;41:237-62.
27. Ditama I, Muktiarti N, Soegijono B. Imidazoline derivatives based on silane functional group as corrosion inhibitor on mild steel. AIP Conf Proc. 2020;2242:020019.
28. Muktiarti N, Ditama I, Soegijono B. Characterization of imidazoline derivates synthesized from soybean oil fatty acids as corrosion inhibitors on mild steel. AIP Conf Proc. 2020;2242:020023.
29. Yotapan N, Sriplai N, Ruengsangtongkul S, Sombatmankhong K. Imidazoline as a volatile corrosion inhibitor for mitigation of top-and bottom-of-the-line CO2 corrosion in carbon steel pipelines. Langmuir. 2024;40:11888-902.
30. Talukdar A, Ganguly D, Roy S, Das N, Sarkar D. Structural evolution and translational potential for agonists and antagonists of endosomal toll-like receptors. J Med Chem. 2021;64:8010-8041.
31. Özbalcı Fİ, Gürbüz N. The Synergistic Effects of LRRK1 Inhibition and TLR4 Activation on Apoptotic, Inflammatory Parameters and α-Synuclein Expression in HMC-3 and SH-SY5Y Cell Lines. TDFD. 2026;15(1):88-96. doi:10.46810/tdfd.1817236.
32. Darendelioğlu E, Sağ S, Caglayan C. Assessment of morin hydrate as a renal protective agent in rats subjected to methotrexate-induced nephrotoxicity. Drug Chem Toxicol. 2025a;48:172-9.
33. Kandemir Ö, Küçükler S, Özdemir S, Dalkılıç E, Sağ S, Caglayan C, et al. Protective effects of quercetin against vincristine‐induced nephrotoxicity via modulation of oxidative stress, inflammation, and apoptosis in rats. J Biochem Mol Toxicol. 2025;39:e70516.
34. Darendelioglu E, Caglayan C, Küçükler S, Bayav İ, Kandemir FM, Ayna A, Sağ S, et al. 18β-glycyrrhetinic acid mitigates bisphenol A-induced liver and renal damage: inhibition of TNF-α/NF-κB/p38-MAPK, JAK1/STAT1 pathways, oxidative stress and apoptosis. Food Chem Toxicol. 2025b;196:115218.