Isolation and Structural Characterization of Ursolic Acid from Medicinal Plant Extract and Its In Silico Inhibitory Potential Against Inflammation-Linked Molecular Targets
Abstract
Objectives:The study aimed to isolate, characterize, and evaluate the neuroprotective and anti-inflammatory potential of a phytochemical compound from a medicinal plant. The primary objective was to assess the binding affinity of ursolic acid, a pentacyclic triterpenoid, against Alzheimer's disease-relevant protein targets using molecular docking. This included enzymes known to contribute to neuroinflammation and amyloid-beta processing, which are implicated in Alzheimer's pathology.
Methods:The crude methanolic extract of the plant was subjected to gradient silica gel column chromatography, followed by purification and recrystallization, yielding a single pure compound. The structure of this compound was confirmed as ursolic acid using FTIR, ¹H-NMR, ¹³C-NMR, and mass spectrometry. Molecular docking was then performed using AutoDock Vina to explore the interaction of ursolic acid with three Alzheimer's-linked targets:
- Glycogen Synthase Kinase-3β (GSK-3β; PDB ID: 1H8F)
- Angiotensin-Converting Enzyme (ACE; PDB ID: 1O86)
- TNF-α Converting Enzyme (TACE; PDB ID: 3LOT)
These proteins were prepared by removing heteroatoms and adding polar hydrogens. Ursolic acid was modeled and energy-minimized using ChemSketch and UCSF Chimera. Binding interactions were visualized using Discovery Studio.
Result:Ursolic acid exhibited favorable binding affinities to all three protein targets involved in neuroinflammation and Alzheimer's progression. The docking results demonstrated stable hydrogen bonding and hydrophobic interactions at the active sites of GSK-3β (linked to tau phosphorylation), ACE (implicated in neurovascular dysfunction), and TACE (key in neuroinflammation via TNF-α activation). The strongest binding affinity was observed with TACE, indicating ursolic acid’s potential in modulating inflammatory cytokine release. These findings suggest that ursolic acid could attenuate both amyloid and inflammatory pathways in Alzheimer's pathology.
Conclusion:This multi-approach study confirmed the identity of ursolic acid and validated its multi-target inhibitory potential against key Alzheimer’s disease-related enzymes. The compound's strong binding affinity, particularly toward TACE and GSK-3β, supports its possible use in modulating neuroinflammation, amyloid cascade, and tau hyperphosphorylation, which are hallmarks of Alzheimer's disease. The study underscores the relevance of plant-derived compounds like ursolic acid in neurodegenerative drug discovery, and supports molecular docking as a predictive tool for evaluating neuroprotective leads
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Kofi Annan, Nora Jackson, Rita A. Dickson, George H. Sam" Gustav Komlaga, Acaricidal effect of an isolate from Hoslundiaoppositavahl against Amblyomma variegatum (Acari: Ixodidae), Pharmacognosy Research 2011,3, 185.
Liu J. Pharmacology of oleanolic acid and ursolic acid. J Ethnopharmacol. 1995;49(2):57-68.
Ikeda Y, Murakami A, Ohigashi H. Ursolic acid: an anti- and pro-inflammatory triterpenoid. Mol Nutr Food Res. 2008;52(1):26-42.
Ríos JL, Francini F. Inflammatory properties of triterpenoids. Phytother Res. 2005;19(9):703-12.
Shanmugam MK, Dai X, Kumar AP, Tan BK, Sethi G, Bishayee A. Ursolic acid in cancer prevention and treatment: molecular targets, pharmacokinetics and clinical studies. BiochemPharmacol. 2013;85(11):1579-87.
Sampath C, Rashid MR, Sang S, Ahmedna M. Bioactive phytochemicals in anti-inflammatory herbs. Food Funct. 2018;9(11):5431-63.
Satyanarayana T, Manohar RD. Ursolic acid–a versatile pentacyclic triterpenoid: pharmacological and analytical aspects. J Pharm Sci Res. 2015;7(8):248-54.
Rakesh K, Anil M, Megha G. Role of GSK-3 in inflammation and its inhibitors as future therapeutic targets. Inflamm Res. 2020;69(3):203-16.
Bernstein KE, Khan Z, Ghosh S, et al. ACE and ACE2: their role in inflammation and fibrosis. Curr Opin Nephrol Hypertens. 2018;27(3):203-10.
Garton KJ, Gough PJ, Blobel CP, et al. Tumor necrosis factor-alpha converting enzyme (TACE/ADAM17) in inflammation and cell signaling. J Biol Chem. 2001;276(43):40241-7.
Morris GM, Huey R, Lindstrom W, et al. AutoDock4 and AutoDockTools4: automated docking with selective receptor flexibility. J Comput Chem. 2009;30(16):2785-91.
Trott O, Olson AJ. AutoDock Vina: improving the speed and accuracy of docking with a new scoring function. J Comput Chem. 2010;31(2):455-61.
Meng XY, Zhang HX, Mezei M, Cui M. Molecular docking: a powerful approach for structure-based drug discovery. Curr Comput Aided Drug Des. 2011;7(2):146-57.
Pettersen EF, Goddard TD, Huang CC, et al. UCSF Chimera–a visualization system for exploratory research and analysis. J Comput Chem. 2004;25(13):1605-12.
Biovia DS. Discovery Studio Modeling Environment, Release 4.1. San Diego: Dassault Systèmes; 2015.
Jones G, Willett P, Glen RC. Molecular recognition of receptor sites using a genetic algorithm with a description of desolvation. J Mol Biol. 1995;245(1):43-53.
Ekins S, Mestres J, Testa B. In silico pharmacology for drug discovery: methods for virtual ligand screening and profiling. Br J Pharmacol. 2007;152(1):9-20.
Berman HM, Westbrook J, Feng Z, et al. The Protein Data Bank. Nucleic Acids Res. 2000;28(1):235-42.
Tautenhahn R, Cho K, Uritboonthai W, Zhu ZJ, Patti GJ, Siuzdak G. An accelerated workflow for untargeted metabolomics using spectral processing and statistical analysis. Anal Chem. 2012;84(11):5071-9.
Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: an overview. ScientificWorldJournal. 2013;2013:162750.
Silva T, Oliveira C, Borges F. Cinnamic acid derivatives as promising anti-inflammatory agents. Curr Med Chem. 2012;19(25):4638-50.
Hu Y, Sun H, Xu F, et al. Ursolic acid derivatives for the treatment of inflammation and cancer: a patent review. Expert Opin Ther Pat. 2020;30(6):423-41.
Yen GC, Duh PD, Tsai HL. Antioxidant and pro-oxidant properties of ascorbic acid and gallic acid. Food Chem. 2002;79(3):307-13.
Panda SK, Swain KC. Phytochemical screening and GC-MS analysis of leaf extract of Ocimum sanctum L. Int J Curr Microbiol Appl Sci. 2011;5(1):1-8.
Trivedi R, Tiwari RK, Parouha YK. Synthesis and characterization of ursolic acid and its derivatives. Indian J Chem B. 2013;52:1324-8.
Yang M, Cheng X, Chen J, et al. Ursolic acid attenuates LPS-induced acute lung injury in mice. Inflamm Res. 2013;62(3):217-26.
Iwata K, Matsuo K, Iida M, et al. Ursolic acid exerts anti-inflammatory effects on skin cells via the MAPK and NF-κB pathways. J Dermatol Sci. 2011;62(3):204-12.
He W, Li Y, Guan H, Xiong Y, Lu Y. Therapeutic potential of ursolic acid against inflammation and cancer. Cancer Lett. 2019;460:1-12.
Singh GB, Singh S. Anti-inflammatory and antimicrobial activities of triterpenoids from Eucalyptus globulus. Fitoterapia. 2005;76(4):329-32.
Bharathi E, Radhika B. Isolation and structural elucidation of ursolic acid from Ocimum sanctum leaves. Int J Pharm Sci Res. 2013;4(7):2586-9.
Zhang Y, Liu Y, Wang T, et al. Ursolic acid inhibits breast cancer progression through repressing cancer stemness via IL-6/STAT3 pathway. Oncotarget. 2017;8(38):70722-33.
Wang X, Li F, Yang X, et al. Anti-inflammatory effects of ursolic acid: a review. Chin J Nat Med. 2020;18(11):803-20.
Yang Y, Liu M, Wu H, et al. Pharmacokinetics and tissue distribution of ursolic acid in rats. J Ethnopharmacol. 2013;146(3):841-6.
Moghadasian MH. Clinical pharmacology of plant sterols. Cardiovasc Ther. 2009;27(2):117-25.
Xu H, Wang Y, Liu C, et al. Ursolic acid modulates NF-κBsignaling in inflammation: a review of preclinical evidence. Front Pharmacol. 2021;12:719693.
Wang C, Wu S, Tu Y, et al. Anti-inflammatory triterpenoids from Rosmarinus officinalis. J Agric Food Chem. 2014;62(19):4517-25.
Fan S, Zhang C, Luo T, et al. Anti-inflammatory effects of ursolic acid via suppression of NF-κB and MAPK pathways in LPS-stimulated RAW 264.7 macrophages. Int Immunopharmacol. 2019;67:223-31.
Rashmi R, Satish V. FTIR and NMR characterization of phytocompounds from medicinal plants. J Pharm Sci Res. 2018;10(4):798-802.
Zhou W, Hu J, Yu L, et al. Targeting GSK3β for treating inflammatory diseases. Front Pharmacol. 2021;12:755034.
Mahato SB, Kundu AP. 13C NMR spectra of pentacyclic triterpenoids – a compilation. Phytochemistry. 1994;37(6):1517-75.
Saini N, Singh D, Sandhu D. FTIR and NMR studies of anti-inflammatory compounds. Int J Pharm Sci Rev Res. 2015;30(2):56-61.
Ghosh R, Chakraborty R, Raychaudhuri U, et al. Role of phytochemicals in cancer chemoprevention. J Food Sci Technol. 2011;48(5):495-505.
Patil S, Patil V, Kadam V. Molecular docking: a review. World J Pharm Res. 2015;4(7):1481-96.
Zhou L, He H, Liu J, et al. Anti-inflammatory and anti-apoptotic effects of ursolic acid on LPS-induced acute kidney injury. Int Immunopharmacol. 2020;89:107040.
Nagao T, Okabe H, Arai I, et al. Search for naturally occurring substances to prevent cancer–anti-tumor promoting activity of triterpenoids. Cancer Lett. 1989;48(2):179-85.
Xu W, Chen X, Shen Y. Anti-inflammatory activity of natural pentacyclic triterpenes. Chin J Nat Med. 2015;13(1):25-35.
Prabhakar PK, Doble M. A target based therapeutic approach towards diabetes mellitus using medicinal plants. Curr Diabetes Rev. 2008;4(4):291-308.
Kim SH, Lee SE, Oh H, et al. Anti-inflammatory and antinociceptive effects of ursolic acid isolated from Corni fructus. Arch Pharm Res. 2011;34(12):2033-41.
Zhao J, Liu J, Cheng H, et al. Ursolic acid exerts neuroprotective effects via Nrf2 signaling. Front Pharmacol. 2020;11:558034.
Aguilar JL, Rojas P, Marcelo A, et al. Anti-inflammatory activity of herbal extracts in macrophages. J Ethnopharmacol. 2002;81(2):271-6.
Lam PY, Jadhav PK. Bioactive natural products as leads in drug discovery. J Med Chem. 2014;57(19):7873-95
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