CK2 and CK1 phosphorylation assays are carried out at 37°C in the presence of increasing amounts of each inhibitor (Ellagic acid) in a final volume of 25 μL containing 50 mM, Tris-HCl pH 7.5, 100 mM NaCl, 12 mM MgCl₂, 0.02 mM [³³P-ATP] (500-1000 cpm/pmol), unless otherwise indicated. The phosphorylatable substrates are the synthetic peptide substrate RRRADDSDDDDD (100 μM) and RRKHAAIGDDDDAYSITA (200 μM) for CK2 and CK1, respectively. Reaction started with the addition of the kinase and is stopped after 10 min. by addition of 5 μL of 0.5 M orthophosphoric acid before spotting aliquots onto phosphocellulose filters. Filters are washed in 75 mM phosphoric acid substrate following SDS-PAGE of the radiolabeled samples. DYRK1A, assayed on the peptide RRRFRPASPLRGPPK, and tyrosine kinase activities are determined^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

ALCL cell viability is measured by MTT assay. Briefly, 0.1 × 10⁵ cells are seeded onto 96-well microculture plates 12 hrs before adding ellagic acid. The cells are grown in 200 μL of complete RPMI-1640 medium, under standard tissue-culture conditions, in the presence or absence of the drug (Ellagic acid) for 48 hours. Twenty μL of MTT solution (5 mg/mL) are then added to the cell suspension for 4h. The intracellular formazan crystals are dissolved with 150 μL of DMSO and optical density, measured on a spectrophotometer at 540 nm, represents the mean (± SD) of triplicate cultures^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Fifty male adult Sprague-Dawley rats are divided randomly into five groups as follow: Group (1) receives corn oil orally as a vehicle and served as normal control. Group (2) receives doxorubicin (DOX) injection (5 mg/kg, i.p.) twice a week for 14 days. Group (3) receives Ellagic acid (10 mg/kg, p.o.; daily) for 14 days and DOX (5 mg/kg, i.p.) twice a week for 14 days. Group (4) receives rosmarinic acid (RA; 75 mg/kg, p.o.; daily) for 14 days and DOX (5 mg/kg, i.p.) twice a week for 14 days. Group (5) receives Ellagic acid (10 mg/kg, p.o.; daily) with RA (75 mg/kg, p.o.; daily) for 14 days and DOX injection (5 mg/kg, i.p.) twice a week for 14 days^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Cozza G, et al. Identification of ellagic acid as potent inhibitor of protein kinase CK2: a successful example of a virtual screening application. J Med Chem. 2006 Apr 20;49(8):2363-6.  [Content Brief]

  [2]. Rizk HA, et al. Prophylactic effects of ellagic acid and rosmarinic acid on doxorubicin-induced neurotoxicity in rats. J Biochem Mol Toxicol. 2017 Dec;31(12).  [Content Brief]

  [3]. Ahire V, et al. Ellagic Acid Enhances Apoptotic Sensitivity of Breast Cancer Cells to γ-Radiation. Nutr Cancer. 2017 Aug-Sep;69(6):904-910.  [Content Brief]

  [4]. Ma CH, et al. Discovery of ellagic acid as a competitive inhibitor of Src homology phosphotyrosyl phosphatase 2 (SHP2) for cancer treatment: In vitro and in silico study. Int J Biol Macromol. 2023 Nov 5;254(Pt 2):127845.  [Content Brief]