{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T05:02:51Z","timestamp":1780549371879,"version":"3.54.1"},"reference-count":30,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T00:00:00Z","timestamp":1780444800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001459","name":"Deputyship for Research & Innovation, Ministry of Education in Saudi Arabia","doi-asserted-by":"publisher","award":["0071-1446-S"],"award-info":[{"award-number":["0071-1446-S"]}],"id":[{"id":"10.13039\/501100001459","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Life"],"abstract":"<jats:p>Background:\u00a0Salvadora persica (S. persica), commonly known as \u2018Miswak,\u2019 has been used in ethnotraditional applications since ancient times. This study was formulated to examine bioactive phenolics and flavonoids from the hydroethanolic extract of S. persica bark, anticancer activity, and in silico binding interaction analysis with key therapeutic targets of triple-negative breast cancer (TNBC) cells. Methods: UHPLC was used to identify the phytochemicals in S. persica bark extract. Cell death was analyzed by MTT assay in TNBC MDA-MB-231 and MDA-MB-468 cells. Moreover, cellular apoptosis, ROS generation, MMP, and cell cycle checkpoints were also carried out. AutoDock Tools 1.5.7 and PyRx 0.8 tools were used for molecular binding interaction analysis. Results: Phytochemical analysis revealed the presence of total phenolic and total flavonoid content of 26.90 \u00b1 0.46 \u03bcg GAE\/mg and 54.51 \u00b1 0.42 \u03bcg QE\/mg of bark extract, respectively. UHPLC analysis confirmed the presence of fumaric acid, chlorogenic acid, rutin, and quercetin in the extract. S. persica significantly reduced cell viability of MDA-MB-231 and MDA-MB-468 cells with an IC50 value of 144 and 128 \u03bcg\/mL, respectively. S. persica extract elevated ROS generation, loss of MMP, late apoptosis induction, and G2\/M-phase cell cycle arrest, while it did not show any significant effect against normal kidney Vero cells. Molecular docking studies revealed that rutin showed strong binding affinity towards EGFR with B.E. = \u22129.8 and \u22129.5 Kcal\/mol; FGFR1 with B.E. = \u22127.4 and \u22127.5 Kcal\/mol; FGFR4 with B.E. = \u22127.5 and \u22127.9 Kcal\/mol; and csGRP78 with B.E. = \u22129.0 and \u22129.3 Kcal\/mol, using Autodock Vina and PyRx tools, respectively. SwissADME and drug-likeness analysis confirmed acceptable drug-like characteristics and favorable pharmacokinetic profiles of the identified molecules. Conclusions: This study highlighted the potential of phytochemicals from S. persica bark as promising compounds for the development of novel anticancer therapeutics.<\/jats:p>","DOI":"10.3390\/life16060943","type":"journal-article","created":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T03:37:29Z","timestamp":1780544249000},"page":"943","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Bioactive Profiling and Evaluation of Anti-Proliferative Potential of Salvadora persica Bark Extract in Triple-Negative Breast Cancer Cells: An In Vitro and Computational Analysis"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3956-1150","authenticated-orcid":false,"given":"Abrar","family":"Turki","sequence":"first","affiliation":[{"name":"Clinical Nutrition Department, College of Applied Medical Sciences, University of Hafr Al Batin, Hafr Al Batin 39524, Eastern Province, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6009-5222","authenticated-orcid":false,"given":"Md. Abul","family":"Barkat","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutics, College of Pharmacy, University of Hafr Al Batin, Hafr Al Batin 39524, Eastern Province, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3082-9368","authenticated-orcid":false,"given":"Yasmin Basheer","family":"Ahmed","sequence":"additional","affiliation":[{"name":"Clinical Nutrition Department, College of Applied Medical Sciences, University of Hafr Al Batin, Hafr Al Batin 39524, Eastern Province, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0374-1096","authenticated-orcid":false,"given":"Harshita","family":"Barkat","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutics, College of Pharmacy, University of Hafr Al Batin, Hafr Al Batin 39524, Eastern Province, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raghad Rashed","family":"Alotaibi","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutics, College of Pharmacy, University of Hafr Al Batin, Hafr Al Batin 39524, Eastern Province, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shivbrat","family":"Upadhyay","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Era\u2019s Lucknow Medical College and Hospital, Era University, Lucknow 226003, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9632-9102","authenticated-orcid":false,"given":"Juveriya","family":"Israr","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Era\u2019s Lucknow Medical College and Hospital, Era University, Lucknow 226003, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0982-1465","authenticated-orcid":false,"given":"Sahabjada","family":"Siddiqui","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Era\u2019s Lucknow Medical College and Hospital, Era University, Lucknow 226003, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Freihat, O., Sipos, D., and Kovacs, A. (2025). Global burden and projections of breast cancer incidence and mortality to 2050: A comprehensive analysis of GLOBOCAN data. Front. Public Health, 13.","DOI":"10.3389\/fpubh.2025.1622954"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"209","DOI":"10.5306\/wjco.v13.i3.209","article-title":"Breast cancer in India: Present scenario and the challenges ahead","volume":"13","author":"Mehrotra","year":"2022","journal-title":"World J. Clin. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1038\/s41523-022-00468-0","article-title":"Triple negative breast cancer: Pitfalls and progress","volume":"8","author":"Zagami","year":"2022","journal-title":"NPJ Breast Cancer"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"102410","DOI":"10.1016\/j.tranon.2025.102410","article-title":"Therapeutic advances and application of PARP inhibitors in breast cancer","volume":"57","author":"Zhou","year":"2025","journal-title":"Transl. Oncol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Chen, J., Wang, Q., Wu, H., Huang, X., and Cao, C. (2025). Therapies targeting triple-negative breast cancer: A perspective on anti-FGFR. Front. Oncol., 14.","DOI":"10.3389\/fonc.2024.1415820"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"881","DOI":"10.1007\/s10439-024-03673-z","article-title":"Chemotherapy-induced cell-surface GRP78 expression as a prognostic marker for invasiveness of metastatic triple-negative breast cancer","volume":"53","author":"Raiter","year":"2025","journal-title":"Ann. Biomed. Eng."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Choudhari, A.S., Mandave, P.C., Deshpande, M., Ranjekar, P., and Prakash, O. (2020). Phytochemicals in cancer treatment: From preclinical studies to clinical practice. Front. Pharmacol., 10.","DOI":"10.3389\/fphar.2019.01614"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1016\/j.sdentj.2011.12.004","article-title":"A review on miswak (Salvadora persica) and its effect on various aspects of oral health","volume":"24","author":"Halawany","year":"2012","journal-title":"Saudi Dent. J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1016\/j.jep.2017.11.030","article-title":"A review of the traditional and modern uses of Salvadora persica L. (Miswak): Toothbrush tree of Prophet Muhammad","volume":"213","author":"Aumeeruddy","year":"2018","journal-title":"J. Ethnopharmacol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"301","DOI":"10.4103\/1305-7456.178297","article-title":"Role of Salvadora persica chewing stick (Miswak): A natural toothbrush for holistic oral health","volume":"10","author":"Niazi","year":"2016","journal-title":"Eur. J. Dent."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5445291","DOI":"10.1155\/2022\/5445291","article-title":"Flavonoids: A bioactive compound from medicinal plants and its therapeutic applications","volume":"2022","author":"Roy","year":"2022","journal-title":"BioMed Res. Int."},{"key":"ref_12","first-page":"e93847","article-title":"Evaluating the antitumor effect of root extract of Salvadora persica in hepatocarcinoma through the induction of apoptosis via an intrinsic pathway: An in vitro study","volume":"17","author":"Latif","year":"2025","journal-title":"Cureus"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1016\/j.sajb.2025.03.041","article-title":"Unlocking hidden biomarkers: Exploring miswak (Salvadora persica L.) potential in tongue squamous cell carcinoma through UPLC-MS\/MS and multivariate analysis","volume":"180","author":"Ibrahim","year":"2025","journal-title":"S. Afr. J. Bot."},{"key":"ref_14","first-page":"4521951","article-title":"Phytochemical, cytotoxic, and antimicrobial evaluation of the fruits of Salvadora persica L.","volume":"2020","author":"Makeen","year":"2020","journal-title":"J. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"253875","DOI":"10.1155\/2014\/253875","article-title":"Assessment of total phenolic and flavonoid content, antioxidant properties, and yield of aeroponically and conventionally grown leafy vegetables and fruit crops: A comparative study","volume":"2014","author":"Chandra","year":"2014","journal-title":"Evid.-Based Complement. Altern. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1038\/s41598-018-36475-0","article-title":"Cytostatic and anti-tumor potential of Ajwa date pulp against human hepatocellular carcinoma HepG2 cells","volume":"9","author":"Siddiqui","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"B\u0142aszczak, E., Miziak, P., Odrzywolski, A., Baran, M., Gumbarewicz, E., and Stepulak, A. (2025). Triple-negative breast cancer progression and drug resistance in the context of epithelial\u2013mesenchymal transition. Cancers, 17.","DOI":"10.3390\/cancers17020228"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"111847","DOI":"10.1016\/j.biopha.2021.111847","article-title":"The impact of the phytotherapeutic agent quercetin on expression of genes and activity of signaling pathways","volume":"141","author":"Shoorei","year":"2021","journal-title":"Biomed. Pharmacother."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"117481","DOI":"10.1016\/j.lfs.2020.117481","article-title":"Targeting PI3K\/Akt\/mTOR signaling pathway by polyphenols: Implication for cancer therapy","volume":"255","author":"Ekrami","year":"2020","journal-title":"Life Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"9155","DOI":"10.2147\/IJN.S518174","article-title":"Quercetin: A natural ally in combating breast cancer","volume":"20","author":"Wu","year":"2025","journal-title":"Int. J. Nanomed."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"717","DOI":"10.3892\/or.2020.7891","article-title":"Chlorogenic acid induces apoptosis, inhibits metastasis and improves antitumor immunity in breast cancer via the NF-\u03baB signaling pathway","volume":"45","author":"Zeng","year":"2021","journal-title":"Oncol. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1186\/bcr597","article-title":"The future of cytotoxic therapy: Selective cytotoxicity based on biology is the key","volume":"5","author":"Tolcher","year":"2003","journal-title":"Breast Cancer Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1038\/s41571-020-0341-y","article-title":"Targeting apoptosis in cancer therapy","volume":"17","author":"Carneiro","year":"2020","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e70107","DOI":"10.1002\/ddr.70107","article-title":"Reactive oxygen species (ROS) and their profound influence on regulating diverse aspects of cancer: A concise review","volume":"86","author":"Tiwari","year":"2025","journal-title":"Drug Dev. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"9051542","DOI":"10.1155\/2019\/9051542","article-title":"ROS-mediated cancer cell killing through dietary phytochemicals","volume":"2019","author":"NavaneethaKrishnan","year":"2019","journal-title":"Oxid. Med. Cell. Longev."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1038\/s41392-024-02080-z","article-title":"Cyclin-dependent protein kinases and cell cycle regulation in biology and disease","volume":"10","author":"Pellarin","year":"2025","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1529","DOI":"10.4155\/fmc.15.80","article-title":"Role of GRP78 in promoting therapeutic-resistant breast cancer","volume":"7","author":"Cook","year":"2015","journal-title":"Future Med. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2593","DOI":"10.1016\/j.apsb.2025.03.004","article-title":"USP51\/GRP78\/ABCB1 axis confers chemoresistance through decreasing doxorubicin accumulation in triple-negative breast cancer cells","volume":"15","author":"Ou","year":"2025","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Tubtimsri, S., Chuenbarn, T., and Manmuan, S. (2025). Quercetin triggers cell apoptosis-associated ROS-mediated cell death and induces S and G2\/M-phase cell cycle arrest in KON oral cancer cells. BMC Complement. Med. Ther., 25.","DOI":"10.1186\/s12906-025-04782-5"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Khan, F., Pandey, P., Jha, N.K., Khalid, M., and Ojha, S. (2021). Rutin mediated apoptotic cell death in Caski cervical cancer cells via Notch-1 and Hes-1 downregulation. Life, 11.","DOI":"10.3390\/life11080761"}],"container-title":["Life"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-1729\/16\/6\/943\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T04:15:28Z","timestamp":1780546528000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-1729\/16\/6\/943"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,3]]},"references-count":30,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["life16060943"],"URL":"https:\/\/doi.org\/10.3390\/life16060943","relation":{},"ISSN":["2075-1729"],"issn-type":[{"value":"2075-1729","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,3]]}}}