Molecular docking analysis of Citrus aurantifolia peel bioactive compounds against fshr, androgen receptor, and PCSK4 in hyperlipidemia-associated male reproductive dysfunction

Roma Sitio* -  Department of Nursing, Polytechnic of Health Ministry of Health, Aceh Besar 23231, Aceh, Indonesia
Dewi Marianthi -  Department of Nursing, Polytechnic of Health Ministry of Health, Aceh Besar 23231, Aceh, Indonesia
Erlangga Galih Zulva Nugroho -  Department of Nursing, Polytechnic of Health Ministry of Health, Aceh Besar 23231, Aceh, Indonesia
Nur Balqis Maulydia -  Eijkman Research Center for Molecular Biology, National Research and Innovation Agency, Jakarta, Indonesia
Teuku Zulfikar -  Department of Pulmonology and Respiratory Medicine, Faculty of Medicine, Universitas Syiah Kuala, Banda Aceh, Indonesia
Wirda Hayati -  Department of Nursing, Polytechnic of Health Ministry of Health, Aceh Besar 23231, Aceh, Indonesia
DOI : 10.30867/action.v11i3.3264

Hyperlipidemia is a diet-induced metabolic disorder that adversely affects systemic metabolism and reproductive function in males. Plant-derived bioactive compounds possess antioxidant, hypolipidemic, and metabolic regulatory activities. This study aimed to investigate the predicted interactions of bioactive compounds from C. aurantifolia peel with proteins involved in metabolic and reproductive pathways using an in silico molecular-docking approach. This in silico molecular docking study was conducted from January to April 2026 at the National Research and Innovation Agency in Indonesia. Ten bioactive compounds identified by GC–MS analysis of Citrus aurantifolia peel ethanol extract were used as test ligands FSHR (PDB ID: 2AM9), androgen receptor (PDB ID: 1XWD), and PCSK4 (UniProt ID: Q6UW60) were prepared using BIOVIA Discovery Studio 2021. Molecular docking was performed using AutoDock Vina in PyRx, with simvastatin as the reference ligand. The docking results were evaluated using the predicted binding affinities and interaction profiles. Pterin-6- carboxylic acid showed the most favorable predicted docking score for FSHR (−7.2 kcal/mol), whereas m-toluic acid showed the most favorable predicted docking scores for the androgen receptor and PCSK4 (−7.5 and −7.8 kcal/mol, respectively). In conclusion, the selected C. aurantifolia peel compounds showed predicted interactions with the three target proteins; however, molecular docking alone does not establish nutraceutical efficacy, bioavailability, safety, receptor activation, or clinical benefit. However, further experimental validation is required.

Keywords : Molecular docking, Citrus aurantifolia, hyperlipidemia, FSHR, androgen receptor, PCSK4

  1. Adel Mehraban, M. S., Tabatabaei-Malazy, O., Rahimi, R., Daniali, M., Khashayar, P., & Larijani, B. (2021). Targeting dyslipidemia by herbal medicines: A systematic review of meta-analyses. Journal of Ethnopharmacology, 280, Article 114407. https://doi.org/10.1016/j.jep.2021.114407
  2. Al-Mohaissen, M. A., Ignaszewski, M. J., Frohlich, J., & Ignaszewski, A. P. (2016). Statin-associated muscle adverse events: Update for clinicians. Sultan Qaboos University Medical Journal, 16(4), e406–e415. https://doi.org/10.18295/squmj.2016.16.04.002
  3. Alzain, A. A., Elbadwi, F. A., Shoaib, T. H., Sherif, A. E., Osman, W., Ashour, A., Mohamed, G. A., Ibrahim, S. R. M., Roh, E. J., & Hassan, A. H. E. (2024). Integrating computational methods guided the discovery of phytochemicals as potential Pin1 inhibitors for cancer: Pharmacophore modeling, molecular docking, MM-GBSA calculations and molecular dynamics studies. Frontiers in Chemistry, 12, Article 1339891. https://doi.org/10.3389/fchem.2024.1339891
  4. Asmah, N., Suniarti, D. F., Margono, A., Mas’ud, Z. A., & Bachtiar, E. W. (2020). Identification of active compounds in ethyl acetate, chloroform, and N-hexane extracts from peels of Citrus aurantifolia from Maribaya, West Java, Indonesia. Journal of Advanced Pharmaceutical Technology & Research, 11(3), 107–112. https://doi.org/10.4103/japtr.JAPTR_177_19
  5. Chopra, B., & Dhingra, A. K. (2021). Natural products: A lead for drug discovery and development. Phytotherapy Research, 35(9), 4660–4702. https://doi.org/10.1002/ptr.7099
  6. Czech, A., Malik, A., Sosnowska, B., & Domaradzki, P. (2021). Bioactive substances, heavy metals, and antioxidant activity in whole fruit, peel, and pulp of citrus fruits. International Journal of Food Science, 2021, Article 6662259. https://doi.org/10.1155/2021/6662259
  7. Dahril, Aulanni’am, Keumala, V., & Mustafa, A. (2019). Human spermatozoa anti-proprotein convertase subtilisin/kexin type 4 synthesis using New Zealand rabbit for novel immunocontraception in males. Investigative and Clinical Urology, 60(4), 303–311. https://doi.org/10.4111/icu.2019.60.4.303
  8. Durmuş, N., Gülsünoğlu-Konuskan, Z., & Kılıç-Akyılmaz, M. (2024). Recovery, bioactivity, and utilization of bioactive phenolic compounds in citrus peel. Food Science & Nutrition, 12(12), 9974–9997. https://doi.org/10.1002/fsn3.4570
  9. El Fadili, M., Er-Rajy, M., Ali Eltayb, W., Kara, M., Assouguem, A., Saleh, A., Al Kamaly, O., Zarougui, S., & Elhallaoui, M. (2023). In-silico screening based on molecular simulations of 3,4-disubstituted pyrrolidine sulfonamides as selective and competitive GlyT1 inhibitors. Arabian Journal of Chemistry, 16(10), Article 105105. https://doi.org/10.1016/j.arabjc.2023.105105
  10. Fekete, M., Lehoczki, A., Kryczyk-Poprawa, A., Zábó, V., Varga, J. T., Bálint, M., Fazekas-Pongor, V., Csípő, T., Rząsa-Duran, E., & Varga, P. (2025). Functional foods in modern nutrition science: Mechanisms, evidence, and public health implications. Nutrients, 17(13), Article 2153. https://doi.org/10.3390/nu17132153
  11. Fitri, Y., Akmal, M., Khairan, K., & Helmi, T. Z. (2026). Male fertility suppression by Piper nigrum: Insights from in vivo and in silico analysis. Journal of Human, Earth, and Future, 7(1), 81–97. https://doi.org/10.28991/HEF-2026-07-01-05
  12. Funes, A. K., Simón, L., Colombo, R., Avena, M. V., Monclús, M., Crescitelli, J., Cabrillana, M. E., Conte, M. I., Cayado, N., Boarelli, P., Fornés, M. W., & Saez Lancellotti, T. E. (2021). Impact of high fat diet on the sterol regulatory element-binding protein 2 cholesterol pathway in the testicle. Molecular Human Reproduction, 27(5), Article gaab023. https://doi.org/10.1093/molehr/gaab023
  13. Galovičová, L., Borotová, P., Vukovic, N. L., Vukic, M., Kunová, S., Hanus, P., Kowalczewski, P. Ł., Bakay, L., & Kačániová, M. (2022). The potential use of Citrus aurantifolia L. essential oils for decay control, quality preservation of agricultural products, and anti-insect activity. Agronomy, 12(3), Article 735. https://doi.org/10.3390/agronomy12030735
  14. García-Fernández-Bravo, I., Torres-Do-Rego, A., López-Farré, A., Galeano-Valle, F., Demelo-Rodriguez, P., & Alvarez-Sala-Walther, L. A. (2022). Undertreatment or overtreatment with statins: Where are we? Frontiers in Cardiovascular Medicine, 9, Article 808712. https://doi.org/10.3389/fcvm.2022.808712
  15. Granato, D., Barba, F. J., Bursać Kovačević, D., Lorenzo, J. M., Cruz, A. G., & Putnik, P. (2020). Functional foods: Product development, technological trends, efficacy testing, and safety. Annual Review of Food Science and Technology, 11, 93–118. https://doi.org/10.1146/annurev-food-032519-051708
  16. Hamad Zubi, Z. B., & Hamad Alfarisi, H. A. (2021). Hyperlipidemia and male infertility. Egyptian Journal of Basic and Applied Sciences, 8(1), 385–396. https://doi.org/10.1080/2314808X.2021.1977080
  17. Indriyani, N. N., & Susanto, A. (2025). Isolasi dan karakterisasi minyak asiri jeruk nipis menggunakan GC-MS dan aktivitasnya terhadap bakteri patogen kulit. OBAT: Jurnal Riset Ilmu Farmasi dan Kesehatan, 3(1), 36–45. https://doi.org/10.61132/obat.v3i1.925
  18. Khutami, C., Sumiwi, S. A., Khairul Ikram, N. K., & Muchtaridi, M. (2022). The effects of antioxidants from natural products on obesity, dyslipidemia, diabetes and their molecular signaling mechanism. International Journal of Molecular Sciences, 23(4), Article 2056. https://doi.org/10.3390/ijms23042056
  19. Mannucci, A., Argento, F. R., Fini, E., Coccia, M. E., Taddei, N., Becatti, M., & Fiorillo, C. (2022). The impact of oxidative stress in male infertility. Frontiers in Molecular Biosciences, 8, Article 799294. https://doi.org/10.3389/fmolb.2021.799294
  20. Matsumoto, T., Koike, M., Arai, C., Kitagawa, T., & Inoue, E. (2018). Chemical structures and antimutagenic effects of unusual oximes from the peels of Citrus limon. Phytochemistry Letters, 25, 118–121. https://doi.org/10.1016/j.phytol.2018.04.016
  21. Maulydia, N. B., Khairan, K., Tallei, T. E., Estevam, E. C., Patwekar, M., Fauzi, F. M., & Idroes, R. (2023). GC-MS analysis reveals unique chemical composition of Blumea balsamifera (L.) DC in Ie-Jue geothermal area. Grimsa Journal of Science Engineering and Technology, 1(1), 9–16. https://doi.org/10.61975/gjset.v1i1.6
  22. Maulydia, N. B., Khairan, K., Tallei, T. E., & Idroes, R. (2025). Metabolomic profiling and anti-inflammatory activity of Blumea balsamifera: Insights from in silico and in vitro studies. Pharmacia, 72, Article e146995. https://doi.org/10.3897/pharmacia.72.e146995
  23. Maulydia, N. B., Wibowo, S., Siregar, J. E., Idroes, R., Syahputra, R. A., & Situmorang, P. C. (2026). Computational insights into the anti-inflammatory potential of Ocimum americanum phytochemicals in malaria-associated cytokine dysregulation. Chemical Methodologies, 10(5), 566–585. https://doi.org/10.48309/chemm.2026.570365.2078
  24. Oduwole, O. O., Huhtaniemi, I. T., & Misrahi, M. (2021). The roles of luteinizing hormone, follicle-stimulating hormone and testosterone in spermatogenesis and folliculogenesis revisited. International Journal of Molecular Sciences, 22(23), Article 12735. https://doi.org/10.3390/ijms222312735
  25. Omolaoye, T. S., Halabi, M. O., Mubarak, M., Cyril, A. C., Duvuru, R., Radhakrishnan, R., & Du Plessis, S. S. (2022). Statins and male fertility: Is there a cause for concern? Toxics, 10(10), Article 627. https://doi.org/10.3390/toxics10100627
  26. Ouattara-Soro, F. S., Acray-Zengbe, P., Febry, K. Y. J., & Abizi, G. (2021). Study of the analgesic effect of the aqueous extract of the leaves of Citrus aurantifolia (Rutaceae) in mice. GSC Biological and Pharmaceutical Sciences, 14(3), 207–214. https://doi.org/10.30574/gscbps.2021.14.3.0072
  27. Pinzi, L., & Rastelli, G. (2019). Molecular docking: Shifting paradigms in drug discovery. International Journal of Molecular Sciences, 20(18), Article 4331. https://doi.org/10.3390/ijms20184331
  28. Quintieri, L., Caputo, L., & Nicolotti, O. (2024). Recent advances in the discovery of novel drugs on natural molecules. Biomedicines, 12(6), Article 1254. https://doi.org/10.3390/biomedicines12061254
  29. Rachmawati, R., Idroes, R., Suhartono, E., Maulydia, N. B., & Darusman, D. (2022). In silico and in vitro analysis of Tacca tubers (Tacca leontopetaloides) from Banyak Island, Aceh Singkil Regency, Indonesia, as antihypercholesterolemia agents. Molecules, 27(23), Article 8605. https://doi.org/10.3390/molecules27238605
  30. Rahayu, V. G., & Hanizar, E. (2021). The effect of lemon (Citrus limon) extracts on the quantity and quality of mice (Mus musculus) sperm. Elkawnie, 7(2), 300–316. https://doi.org/10.22373/ekw.v7i2.9389
  31. Rose, M., Duhamel, M., Rodet, F., & Salzet, M. (2021). The role of proprotein convertases in the regulation of the function of immune cells in the oncoimmune response. Frontiers in Immunology, 12, Article 667850. https://doi.org/10.3389/fimmu.2021.667850
  32. Santini, A., Cammarata, S. M., Capone, G., Ianaro, A., Tenore, G. C., Pani, L., & Novellino, E. (2018). Nutraceuticals: Opening the debate for a regulatory framework. British Journal of Clinical Pharmacology, 84(4), 659–672. https://doi.org/10.1111/bcp.13496
  33. Sitio, R., Akmal, M., Marlina, M., & Gholib, G. (2024). Investigating ethanolic extract from Acehnese lime (Citrus aurantifolia) peel as potential anti-hypercholesterolemia agent. Journal of Human, Earth, and Future, 5(3), 348–365. https://doi.org/10.28991/HEF-2024-05-03-04
  34. Sutrio, S., Nugroho, A., Wahyuni, E. S., & Putri, S. (2026). Association of dietary intake and physical activity with hypercholesterolemia among Prolanis participants. AcTion: Aceh Nutrition Journal, 11(1), 205–214. https://doi.org/10.30867/action.v11i1.3063
  35. Tanveer, A., Akram, K., Farooq, U., Hayat, Z., & Shafi, A. (2017). Management of diabetic complications through fruit flavonoids as a natural remedy. Critical Reviews in Food Science and Nutrition, 57(7), 1411–1422. https://doi.org/10.1080/10408398.2014.1000482
  36. Tareq, A. M., Mahmud, M. H., Billah, M. M., Hasan, M. N., Jahan, S., Hossain, M. M., Chy, F. J., Uddin, M. G., Emran, T. B., & Sayeed, M. A. (2022). Fast-food and obesity: Status among the young adult population in Bangladesh. Narra J, 2(3), Article e86. https://doi.org/10.52225/narraj.v2i3.86
  37. Wang, J. M., Li, Z. F., Yang, W. X., & Tan, F. Q. (2022). Follicle-stimulating hormone signaling in Sertoli cells: A licence to the early stages of spermatogenesis. Reproductive Biology and Endocrinology, 20(1), Article 97. https://doi.org/10.1186/s12958-022-00971-w
  38. World Health Organization. (2025, November 28). Infertility. https://www.who.int/news-room/fact-sheets/detail/infertility
  39. Zubi, Z. B. H., & Alfarisi, H. A. H. (2021). Hyperlipidemia and male infertility. Egyptian Journal of Basic and Applied Sciences, 8(1), 385–396. https://doi.org/10.1080/2314808X.2021.1977080

Open Access Copyright (c) 2026 Roma Sitio, Dewi Marianthi, Erlangga Galih Zulva Nugroho, Nur Balqis Maulydia, Teuku Zulfikar, Wirda Hayati
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