HPTLC Identification and Quantification of 2,5-Di-tert-butyl-1,4-benzoquinone in Flax Microgreen Extracts: Exploring its Anticancer Potential Against Prostate Cancer

Abstract

Prostate cancer is a hereditary disease, the second deadliest and most prevalent malignancy among men. In pharmacotherapy, natural compounds and their structural analogs have been used for cancer treatment. Several studies have revealed the potentiality of Linum usitatissimum in various cancer treatments. This study aimed to identify and quantify 2,5-Di-tert-butyl-1,4-benzoquinone (2,5-DTBQ) from methanolic extract of flax microgreens (MEFM) using GC-MS and HPTLC, and to assess its inhibitory activity against prostate cancer. 2,5-DTBQ was identified and quantified by GC-MS and HPTLC analysis. The docking simulation had been carried out in PyRx 0.8 software. Toxicity studies were performed using ADMETlab 3.0 and ProTox 3.0 prediction tools, respectively. The cytotoxic effects and induction of apoptotic cell death by 2,5-DTBQ on PC-3 cell lines were assessed by MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and annexin V apoptosis assays, respectively. HPTLC fingerprint has proven that there is a high content of 2,5-DTBQ in MEFM. The docking result revealed that the 2,5-DTBQ compound exhibited the highest binding affinities and favorable interactions against prostate cancer target proteins. The ADME/T and toxicity prediction studies demonstrated that the 2,5-DTBQ compound had low toxicity and specific metabolic characteristics. 2,5-DTBQ had exhibited mild growth inhibition against PC-3 cell lines with the IC₅₀ value of 947.03 μg/ml. These findings had proven that 2,5-DTBQ served as a novel inhibitor for prostate cancer treatment, but it can further be confirmed via in vivo studies.

Keywords: 2,5-DTBQ; ADME; anti-prostate cancer; flax microgreens; GC-MS; HPTLC; molecular docking.