EFFICIENT AND SENSITIVE ANALYSIS OF BIOMOLECULAR INTERACTIONS BETWEEN PALBOCICLIB AND HUMAN SERUM ALBUMIN USING FLUORESCENCE QUENCHING MECHANISM

Öz

Objective: In this research, the interaction between Palbociclib (PC) and Human Serum Albumin (HSA) was examined using fluorescence spectroscopy, and the quenching mechanism was elucidated. Material and Method: HSA and PC stock solutions were prepared using 0.1 M pH 7.4 phosphate buffer solution and DMSO, respectively. In the protein-drug interaction experiment, the HSA concentration was fixed at 20 µM, and drug concentrations for the PC-HSA system were used between 0 and 40 µM. Result and Discussion: The interaction between PC and HSA was investigated at various temperatures using spectroscopic techniques. To comprehend the interaction process, experiments were carried out at different physiological pH levels and temperatures. Other parameters, including the number of binding sites, binding constants, and quenching constants, were calculated using the proper formulas. The van't Hoff equation was used to determine thermodynamic parameters such as entropy, Gibbs, and enthalpy at different temperatures.

Anahtar Kelimeler

• Drug-protein interaction
• fluorescence spectroscopy
• human serum albumin
• palbociclib
• quenching mechanism

PALBOSİKLİB VE İNSAN SERUM ALBÜMİN ARASINDAKI BİYOMOLEKÜLER ETKİLEŞİMLERİN FLORESAN SÖNÜMLEME MEKANİZMASI KULLANILARAK ETKİLİ VE HASSAS BİR ŞEKİLDE ANALİZ EDİLMESİ

Öz

Amaç: Bu çalışmada, Palbosiklib (PC) ile İnsan Serum Albümini (HSA) arasındaki etkileşim floresan spektroskopisi kullanılarak incelendi ve sönümleme mekanizması aydınlatıldı. Gereç ve Yöntem: HSA ve PC stok çözeltileri sırasıyla 0.1 M pH 7.4 fosfat tampon çözeltisi ve DMSO kullanılarak hazırlandı. Protein-ilaç etkileşimi deneyinde, HSA konsantrasyonu 20 µM'de sabitlendi ve PC-HSA sistemi için ilaç konsantrasyonları 0 ila 40 µM arasında kullanıldı. Sonuç ve Tartışma: PC ve HSA arasındaki etkileşim, spektroskopik teknikler kullanılarak çeşitli sıcaklıklarda incelenmiştir. Etkileşim sürecini anlamak için farklı fizyolojik pH seviyelerinde ve sıcaklıklarda deneyler gerçekleştirilmiştir. Bağlanma bölgesi sayısı, bağlanma sabitleri ve söndürme sabitleri gibi diğer parametreler, uygun formüller kullanılarak bulunmuştur. van't Hoff denklemi, farklı sıcaklıklarda entropi, Gibbs ve entalpi gibi termodinamik parametrelerini belirlemek için kullanıldı.

Anahtar Kelimeler

• Floresan spektroskopisi
• ilaç-protein etkileşimi
• insan serum albümini
• palbosiklib
• sönümleme mekanizması

Kaynakça

1. 1. Khan, M.I., Nadeem, I., Majid, A., et al. (2021). Adsorption mechanism of Palbociclib anticancer drug on two different functionalized nanotubes as a drug delivery vehicle: A first principle’s study. Applied Surface Science, 546, 149129. [CrossRef]
2. 2. Wang, Y., Wang, F., Shen, Y., et al. (2018). Tumor-specific disintegratable nanohybrids containing ultrasmall inorganic nanoparticles: From design and improved properties to cancer applications. Materials Horizons, 5(2), 184-205. [CrossRef]
3. 3. Chu, C., Lyu, X., Wang, Z., et al. (2020). Cocktail polyprodrug nanoparticles concurrently release cisplatin and peroxynitrite-generating nitric oxide in cisplatin-resistant cancers. Chemical Engineering Journal, 402, 126125. [CrossRef]
4. 4. Inglot, T.W. (2025). Exploring pitolisant binding to human serum albumin: Insights from multi-spectroscopic, molecular docking and molecular dynamics studies. Journal of Molecular Structure, 1341, 142698. [CrossRef]
5. 5. Zsila, F. (2013). Subdomain IB is the third major drug binding region of human serum albumin: Toward the three-sites model. Molecular Pharmaceutics, 10(5), 1668-1682. [CrossRef]
6. 6. Colmenarejo, G. (2003). In silico prediction of drug-binding strengths to human serum albumin. In Medicinal Research Reviews. 23 (3), 275-301. [CrossRef]
7. 7. Ghuman, J., Zunszain, P.A., Petitpas, I., et al. (2005). Structural basis of the drug-binding specificity of human serum albumin. Journal of Molecular Biology, 353(1), 38-52. [CrossRef]
8. 8. Calucică, D.M., Manda, C.V., Găman, A.M., et al. (2022). Development of a SPE-LC-MS method for the Quantitation of palbociclib and abemaciclib in human plasma. Molecules, 27(23), 8604. [CrossRef]

9. 9. Mlinarić, Z., Turković, L., Begović, I., et al. (2022). Rapid capillary electrophoresis method for simultaneous determination of abemaciclib, ribociclib, and palbociclib in pharmaceutical dosage forms: A green approach. Molecules, 27(21), 7603. [CrossRef]
10. 10. Magdy, G., Belal, F., Elmansi, H. (2023). Rapid microwave-assisted synthesis of nitrogen-doped carbon quantum dots as fluorescent nanosensors for the spectrofluorimetric determination of palbociclib: Application for cellular imaging and selective probing in living cancer cells. RSC Advances, 13(7), 4156-4167. [CrossRef]
11. 11. Bilge, S., Dogan Topal, B., Caglayan, M.G., et al. (2022). Human hair rich in pyridinic nitrogen-base DNA biosensor for direct electrochemical monitoring of palbociclib-DNA interaction. Bioelectrochemistry, 148, 108264. [CrossRef]
12. 12. Cheraghi, S., Cetinkaya, A., Kaya, S.I., et al. (2024). Sensitive and selective electrochemical sensor for palbociclib, a highly selective CDK4/6 inhibitor, based on molecularly imprinted polymer. Microchemical Journal, 201, 110689. [CrossRef]
13. 13. Cheraghi, S., Taher, M. A., Karimi-Maleh, H. (2017). A sensitive amplified sensor based on improved carbon paste electrode with 1-methyl-3-octylimidazolium tetrafluoroborate and ZnO/CNTs nanocomposite for differential pulse voltammetric analysis of raloxifene. Applied Surface Science, 420, 882-885. [CrossRef]
14. 14. Bozoǧlan, B.K., Tunç, S., Duman, O. (2014). Investigation of neohesperidin dihydrochalcone binding to human serum albumin by spectroscopic methods. Journal of Luminescence, 155, 198-204. [CrossRef]
15. 15. Xiong, X., He, J., Yang, H., et al. (2017). Investigation on the interaction of antibacterial drug moxifloxacin hydrochloride with human serum albumin using multi-spectroscopic approaches, molecular docking, and dynamical simulation. RSC Adv., 7(77), 48942-48951. [CrossRef]
16. 16. Caruso, Í.P., Vilegas, W., Cristante de Oliveira, L., et al. (2020). Fluorescence spectroscopic and dynamics simulation studies on isoorientin binding with human serum albumin. Spectrochimica Acta - Part A: Molecular and Biomolecular Spectroscopy, 228, 117738. [CrossRef]
17. 17. Li, Y., He, W.Y., Dong, Y.M., et al. (2006). Human serum albumin interaction with formononetin studied using fluorescence anisotropy, FT-IR spectroscopy, and molecular modeling methods. Bioorganic and Medicinal Chemistry, 14(5), 1431-1436. [CrossRef]
18. 18. Cetinkaya, A., Caglayan, M.G., Unal, M.A., et al. (2022). Investigation of pazopanib and human serum albumin interaction using spectroscopic and molecular docking approaches. Analytica, 3(1), 144-160. [CrossRef]
19. 19. Gonzàlez-Jimènez, J., Frutos, G., Cayre, I. (1992). Fluorescence quenching of human serum albumin by xanthines. Biochemical Pharmacology, 44(4), 824-826.[CrossRef]
20. 20. Fan, X.Y., Zhang, Y., Wang, J., et al. (2014). Exploring the interaction between rotenone and human serum albumin. The Journal of Chemical Thermodynamics, 69, 186-192. [CrossRef]
21. 21. Zhang, W., Zhong, H., Zhao, P., et al. (2022). Carbon quantum dot fluorescent probes for food safety detection: Progress, opportunities and challenges. Food Control, 133, 108591. [CrossRef]
22. 22. Tunç, S., Çetinkaya, A., Duman, O. (2013). Spectroscopic investigations of the interactions of tramadol hydrochloride and 5-azacytidine drugs with human serum albumin and human hemoglobin proteins. Journal of Photochemistry and Photobiology B: Biology, 120, 59-65. [CrossRef]