Integrative pharmacological profiling of leaf-derived steroids from Belosynapsis vivipara for Ulcer protection: In silico and in vivo approaches
Abstract
This investigation focused on evaluating the anti-ulcer properties of a steroidal compound extracted from the ethanol-based leaf extract of Belosynapsis vivipara. A combination of polar and nonpolar solvents facilitated the isolation process, and the resulting fractions were characterized using thin-layer chromatography, Fourier-transform infrared spectroscopy, and high-performance liquid chromatography. The extract was administered orally to Wistar albino rats at doses of 250 mg/kg and 500 mg/kg, with ethanol-induced gastric lesions serving as the experimental model. Ranitidine was used as the reference drug for comparison. To support the biological findings, molecular docking and ADMET profiling were conducted against human carbonic anhydrase isoforms I and II. β-sitosterol was identified as a major constituent through column chromatography. The higher dose of the extract significantly improved ulcer-related parameters, including ulcer index, gastric pH, and acidity levels, suggesting a dose-dependent therapeutic effect. Docking analysis revealed notable interactions between β-sitosterol and the target protein (PDB ID: 6g3v), including hydrogen bonding with ALA132 and hydrophobic contacts with ALA135 and LEU131. The compound also met Lipinski’s criteria for drug-likeness. These findings indicate that the ethanol extract of B. vivipara leaves, particularly due to β-sitosterol, holds promise as a natural agent for gastric ulcer management through mucosal protection and acid regulation.
Keywords:
Antiulcer Activity, Belosynapsis vivipara, Beta-Sitosterol, Docking Studies, Isolation
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References
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Das, K. (2025). Microscopic characterization and proximate analysis of Belosynapsis vivipara leaves: A rare plant of Western Ghats region. Vegetos, 1-20. https://doi.org/10.1007/s42535-025-01338-3
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Hamed, S. N. N., Fahmy, N. M., Singab, A. N. B., & Mohamed, R. W. (2022). Anti-ulcer effects of cumin (Cuminum cyminum L.), thyme (Thymus vulgaris L.), and caraway (Carum carvi L.) essential oils on peptic ulcer and ulcerative colitis models in rats. Journal of HerbmedPharmacolog,11(3), 389-400. https://doi.org/10.34172/jhp.2022.45
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Kavade, S., Deokule, S., Lakshminarasimhan, P., Diwakar, P., & Punekar, S. (2012). Belosynapsis vivipara (Dalzell) CEC Fisch. (Commelinaceae), a vulnerable spiderwort, rediscovered after sixteen decades from Maharashtra, India. Journal of Threatened Taxa, 4(6), 2660-2663. https://doi.org/10.11609/JoTT.o2444.2660-3
Kohale, S. P., Kubde, J. A., Hatwar, P. R., Bakal, R. L., &Sapkal, R. N. (2025). Medicinal plants with anti-ulcer properties: A review. International Journal of Herbal Medicine,13(4), 124-130. https://www.doi.org/10.22271/flora.2025.v13.i4b.1006
Kumar, Y. S., Selvam, N. T., Gaidhani, N. S., Lamba, D., Sharma, B. S., Chauhan, V. S., & Srikanth, N. (2025). Acute and sub-chronic oral toxicity study of classical ayurvedic formulation yogarajaguggulu. Research Journal of Pharmacy and Technology, 18(5), 2175-2180. http://dx.doi.org/10.52711/0974-360X.2025.00312
Kurian, T. (2024). Comparative in silico molecular docking analysis of sophocarpine and chelerythrine alkaloids for antiherpes activity. Indian Drugs, 61(6), 74-76. https://doi.org/10.53879/id.61.06.14542
Li, S., Bao, S., Cai, L., Li, B., Sun, Y., & Sun, Y. (2025). Acute and subacute toxicity evaluation of ZhenzhuXiaoji decoction in preclinical models: implications for safe clinical use. Frontiers in Pharmacology, 16, 1554732. https://doi.org/10.3389/fphar.2025.1554732
Loosli, A. W. M., Rhoden, S. A., Selari, P. J. R. G., Freitas, G. F. D., Silva, T. B. D., Ricci, A. P., ... & Almeida, T. T. D. (2024). In silico analysis of the biodiversity of endophytic fungi isolated from the Brazilian Cerrado. Biota Neotropica, 24(2), e20231603. https://doi.org/10.1590/1676-0611-BN-2023-1603
Mesileya, K. T., Onyeka, P. C., Adaramola, I. M., Igbalaye, Q. O., Bodun, D. S., Alao, W. K., ... &Omotuyi, O. I. (2025). Tinospora cordifolia bioactive compounds as a novel sterol 14a-demethylase (CYP51) inhibitor: An in silico study. In Silico Pharmacology, 13(1), 28. https://doi.org/10.1007/s40203-025-00312-w
Miszczuk, E., Bajguz, A., Kiraga, Ł., Crowley, K., &Chłopecka, M. (2024). Phytosterols and the digestive system: A review study from insights into their potential health benefits and safety. Pharmaceuticals, 17(5), 557. https://doi.org/10.3390/ph17050557
Okokon, J. E., Etuk, I. C., Thomas, P. S., Drijfhout, F. P., Claridge, T. D., & Li, W. W. (2022). In vivo antihyperglycaemic and antihyperlipidemic activities and chemical constituents of Solanum anomalum. Biomedicine & Pharmacotherapy, 151, 113153. https://doi.org/10.1016/j.biopha.2022.113153
Patidar, H., Saxena, R., & Chouhan, M. (2021). Evaluation of the anti-diarrheal and anti-ulcer activity of hydro-alcoholic extract of aerial part of Tridax procumbens in experimental rats. Advance Pharmaceutical Journal, 6(3), 65-74. https://doi.org/10.31024/apj.2021.6.3.2
Rashid, K.H., Sharma, S., Dhawan, A., & Sharma, D. (2025). Peptic ulcers in 2025: New insights and modern treatment approaches. International Journal of Pharmacology and Clinical Research, 7(1), 112-117. https://www.doi.org/10.33545/26647613.2025.v7.i1b.61
Rathod, S., Gidnavar, L., Arakeri, M., Nadaf, N., Rathod, N., & Galgali, N. (2025). Investigation of In-Vitro Anti-Ulcer Activity of Whole Plant Methanolic Extract of Ficus Religiosa. International Journal of Pharmaceutical Drug Design, 2(12), 26-39. https://doi.org/10.62896/ijpdd.2.12.02
Ruba, S. Y., Sule, M. I., Sani, Y. M., Sadam, A. A., &Uzairu, M. S. (2024). Isolation and Structural Elucidation of Stigmasterol, β-Sitosterol, and Tripalmitolein from the Leaf of Vitex Chrysocarpa (Lamiaceae). UMYU Scientifica, 3(4), 84-93. https://doi.org/10.56919/usci.2434.008
Sasane, N. A., Gaikar, P. S., Gaikwad, S. L., Valvi, A. K., & Wadhawa, G. C. (2021). Phytochemical analysis, anti-oxidant and anti-inflammatory of Belosynapsis vivipara leaves and roots. International Journal of Botany Studies, 6(4), 138-142. https://www.botanyjournals.com/assets/archives/2021/vol6issue4/6-4-14-258.pdf
Sharma, D. K., Sharma, S. K., Patil, R. D., Lonare, M., & Sharma, A. (2025). Evaluation of antiulcer activity of ethanolic extract of leaf, bark, flower and fruit pulp of Cassia fistula in Wistar rats. Toxicology International, 32(1), 49–56. https://doi.org/10.18311/ti/2025/v32i1/46304
Vani, V., S., S. Nafiya, F., A, H., ……. Saji, S., (2024). In-Silico Design of Thiadiazole Derivatives as Anticonvulsant Agents. Asian Journal of Pharmaceutical Research and Development, 12(4), 85-91. http://dx.doi.org/10.22270/ajprd.v12i4.1445
Yigit, S., Kaci, F. N., Gezer, A., Yayla, M., Duysak, L., Aksu Kilicle, P., ... &Bingol, S. A. (2025). Gastroprotective and antioxidant effects of Ferula plant extract against indomethacin induced gastric ulcer in rats. Food Science & Nutrition, 13(8), e70730. https://doi.org/10.1002/fsn3.70730
Yuenyong, J., Bennett, C., Jiamyangyuen, S., Mahatheeranont, S., &Sookwong, P. (2024). Development of a simultaneous normal-phase HPLC analysis of lignans, tocopherols, phytosterols, and squalene in sesame oil samples. Foods, 13(9), 1368. https://doi.org/10.3390/foods13091368
Dali, N., Chairunnas, A., & Amalia, H. A. M. (2024). Extraction and purification of steroids in the sea urchin (Diadema Setosum Leske) gonads with a three-component solvent system. Biointerface Research in Applied Chemistry, 14(3), 1-19. https://doi.org/10.33263/BRIAC143.066
Das, K. (2013). Wound healing potential of aqueous crude extract of Stevia rebaudiana in mice. Revista Brasileira de Farmacognosia, 23(2), 351-357. https://doi.org/10.1590/S0102-695X2013005000011
Das, K. (2025). Microscopic characterization and proximate analysis of Belosynapsis vivipara leaves: A rare plant of Western Ghats region. Vegetos, 1-20. https://doi.org/10.1007/s42535-025-01338-3
Das, K., &Singirikonda, S. (2023). Elemental impact on antibacterial study of hydroalcoholic leaves extract of Belosynapsis vivipara. Notulae Scientia Biologicae, 15(1), 11409. https://doi.org/10.55779/nsb15111409
Das, K., Dang, R., Sivaraman, G., &Ellath, R. P. (2018). Phytochemical screening for various secondary metabolites, antioxidant, and anthelmintic activity of Cosciniumfenestratum fruit pulp: A new biosource for novel drug discovery. Turkish Journal of Pharmaceutical Sciences, 15(2), 156-165. https://doi.org/10.4274/tjps.54376
Das, P., Nayak, A., Bhavani, K., & Afsar, M. (2023). Molecular modelling technique on interactivity between human carbonic anhydrase 1 and mangiferin for antiulcer activity. Research Journal of Pharmacy and Technology, 16(5), 2465-2469. http://dx.doi.org/10.52711/0974-360X.2023.00406
Ghareeb, M. A., Mohammed, H. S., Aboushousha, T., Lotfy, D. M., El-Shazly, M. A., Sobeh, M., & Taha, E. F. (2024). Ipomoea carnea mitigates ethanol-induced ulcers in irradiated rats via Nrf2/HO− 1 pathway: An in vivo and in silico study. Scientific Reports, 14, 3469. https://doi.org/10.1038/s41598-024-53336-1
Gupta, H., Shakya, M. K., & Sharma, G. K. (2022). Evaluation of Anti-ulcer activity of Polyherbal preparation in Ulcerogenic Wistar rats. Research Journal of Pharmacy and Technology, 15(8), 3471-3474. http://dx.doi.org/10.52711/0974-360X.2022.00581
Hamed, S. N. N., Fahmy, N. M., Singab, A. N. B., & Mohamed, R. W. (2022). Anti-ulcer effects of cumin (Cuminum cyminum L.), thyme (Thymus vulgaris L.), and caraway (Carum carvi L.) essential oils on peptic ulcer and ulcerative colitis models in rats. Journal of HerbmedPharmacolog,11(3), 389-400. https://doi.org/10.34172/jhp.2022.45
Hieu, N. V., Vinh, L. B., Phong, N. V., Cong, P. V., Dat, N. T., Dan, N. V., ... & Tuan Anh, H. L. (2024). Two new steroidal saponins with potential anti-inflammatory effects from the aerial parts of GnetumformosumMarkgr. Plants, 13(15), 2100. https://doi.org/10.3390/plants13152100
Jazeela, D’Souza, U.P., Jyothi, D., & Joshi, H. (2022). Evaluation of antiulcer activity of alcoholic stem extract of Epipremnumaureum (Linden andre) G.S. Bunting. Research Journal of Pharmacy and Technology,15(12), 5779-5784. https://doi.org/10.52711/0974-360X.2022.00975
Kalyani, G., Fathima, A., Venkateswarlu, G., Reddy, D. R. S., & Rasheed, S. H. (2023). Evaluation of antibacterial and antioxidant potential of Melia. azedarach linn. and Psidium guajava linn. leaf extracts. Research Journal of Pharmacy and Technology, 16(7), 3132-3138. http://dx.doi.org/10.52711/0974-360X.2023.00515
Kavade, S., Deokule, S., Lakshminarasimhan, P., Diwakar, P., & Punekar, S. (2012). Belosynapsis vivipara (Dalzell) CEC Fisch. (Commelinaceae), a vulnerable spiderwort, rediscovered after sixteen decades from Maharashtra, India. Journal of Threatened Taxa, 4(6), 2660-2663. https://doi.org/10.11609/JoTT.o2444.2660-3
Kohale, S. P., Kubde, J. A., Hatwar, P. R., Bakal, R. L., &Sapkal, R. N. (2025). Medicinal plants with anti-ulcer properties: A review. International Journal of Herbal Medicine,13(4), 124-130. https://www.doi.org/10.22271/flora.2025.v13.i4b.1006
Kumar, Y. S., Selvam, N. T., Gaidhani, N. S., Lamba, D., Sharma, B. S., Chauhan, V. S., & Srikanth, N. (2025). Acute and sub-chronic oral toxicity study of classical ayurvedic formulation yogarajaguggulu. Research Journal of Pharmacy and Technology, 18(5), 2175-2180. http://dx.doi.org/10.52711/0974-360X.2025.00312
Kurian, T. (2024). Comparative in silico molecular docking analysis of sophocarpine and chelerythrine alkaloids for antiherpes activity. Indian Drugs, 61(6), 74-76. https://doi.org/10.53879/id.61.06.14542
Li, S., Bao, S., Cai, L., Li, B., Sun, Y., & Sun, Y. (2025). Acute and subacute toxicity evaluation of ZhenzhuXiaoji decoction in preclinical models: implications for safe clinical use. Frontiers in Pharmacology, 16, 1554732. https://doi.org/10.3389/fphar.2025.1554732
Loosli, A. W. M., Rhoden, S. A., Selari, P. J. R. G., Freitas, G. F. D., Silva, T. B. D., Ricci, A. P., ... & Almeida, T. T. D. (2024). In silico analysis of the biodiversity of endophytic fungi isolated from the Brazilian Cerrado. Biota Neotropica, 24(2), e20231603. https://doi.org/10.1590/1676-0611-BN-2023-1603
Mesileya, K. T., Onyeka, P. C., Adaramola, I. M., Igbalaye, Q. O., Bodun, D. S., Alao, W. K., ... &Omotuyi, O. I. (2025). Tinospora cordifolia bioactive compounds as a novel sterol 14a-demethylase (CYP51) inhibitor: An in silico study. In Silico Pharmacology, 13(1), 28. https://doi.org/10.1007/s40203-025-00312-w
Miszczuk, E., Bajguz, A., Kiraga, Ł., Crowley, K., &Chłopecka, M. (2024). Phytosterols and the digestive system: A review study from insights into their potential health benefits and safety. Pharmaceuticals, 17(5), 557. https://doi.org/10.3390/ph17050557
Okokon, J. E., Etuk, I. C., Thomas, P. S., Drijfhout, F. P., Claridge, T. D., & Li, W. W. (2022). In vivo antihyperglycaemic and antihyperlipidemic activities and chemical constituents of Solanum anomalum. Biomedicine & Pharmacotherapy, 151, 113153. https://doi.org/10.1016/j.biopha.2022.113153
Patidar, H., Saxena, R., & Chouhan, M. (2021). Evaluation of the anti-diarrheal and anti-ulcer activity of hydro-alcoholic extract of aerial part of Tridax procumbens in experimental rats. Advance Pharmaceutical Journal, 6(3), 65-74. https://doi.org/10.31024/apj.2021.6.3.2
Rashid, K.H., Sharma, S., Dhawan, A., & Sharma, D. (2025). Peptic ulcers in 2025: New insights and modern treatment approaches. International Journal of Pharmacology and Clinical Research, 7(1), 112-117. https://www.doi.org/10.33545/26647613.2025.v7.i1b.61
Rathod, S., Gidnavar, L., Arakeri, M., Nadaf, N., Rathod, N., & Galgali, N. (2025). Investigation of In-Vitro Anti-Ulcer Activity of Whole Plant Methanolic Extract of Ficus Religiosa. International Journal of Pharmaceutical Drug Design, 2(12), 26-39. https://doi.org/10.62896/ijpdd.2.12.02
Ruba, S. Y., Sule, M. I., Sani, Y. M., Sadam, A. A., &Uzairu, M. S. (2024). Isolation and Structural Elucidation of Stigmasterol, β-Sitosterol, and Tripalmitolein from the Leaf of Vitex Chrysocarpa (Lamiaceae). UMYU Scientifica, 3(4), 84-93. https://doi.org/10.56919/usci.2434.008
Sasane, N. A., Gaikar, P. S., Gaikwad, S. L., Valvi, A. K., & Wadhawa, G. C. (2021). Phytochemical analysis, anti-oxidant and anti-inflammatory of Belosynapsis vivipara leaves and roots. International Journal of Botany Studies, 6(4), 138-142. https://www.botanyjournals.com/assets/archives/2021/vol6issue4/6-4-14-258.pdf
Sharma, D. K., Sharma, S. K., Patil, R. D., Lonare, M., & Sharma, A. (2025). Evaluation of antiulcer activity of ethanolic extract of leaf, bark, flower and fruit pulp of Cassia fistula in Wistar rats. Toxicology International, 32(1), 49–56. https://doi.org/10.18311/ti/2025/v32i1/46304
Vani, V., S., S. Nafiya, F., A, H., ……. Saji, S., (2024). In-Silico Design of Thiadiazole Derivatives as Anticonvulsant Agents. Asian Journal of Pharmaceutical Research and Development, 12(4), 85-91. http://dx.doi.org/10.22270/ajprd.v12i4.1445
Yigit, S., Kaci, F. N., Gezer, A., Yayla, M., Duysak, L., Aksu Kilicle, P., ... &Bingol, S. A. (2025). Gastroprotective and antioxidant effects of Ferula plant extract against indomethacin induced gastric ulcer in rats. Food Science & Nutrition, 13(8), e70730. https://doi.org/10.1002/fsn3.70730
Yuenyong, J., Bennett, C., Jiamyangyuen, S., Mahatheeranont, S., &Sookwong, P. (2024). Development of a simultaneous normal-phase HPLC analysis of lignans, tocopherols, phytosterols, and squalene in sesame oil samples. Foods, 13(9), 1368. https://doi.org/10.3390/foods13091368
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Das, K., Das, P., Raja, R., Murthy, L. and Nayak, A. (2026) “Integrative pharmacological profiling of leaf-derived steroids from Belosynapsis vivipara for Ulcer protection: In silico and in vivo approaches”, International Journal of Advancement in Life Sciences Research, 9(3), pp. 11-24. doi: https://doi.org/10.31632/ijalsr.2026.v09i03.002.
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