Novel triterpenoid from Citrus aurantium L. possesses chemopreventive properties against human colon cancer cells.
Potential cancer preventive constituents of sour orange (Citrus aurantium L.) were isolated and identified from EtOAc extract of sour orange. Crude EtOAc extract was purified using silica gel column chromatography to isolate two putative bioactive compounds. The purity of the isolated compounds was analyzed by TLC and HPLC. The structures of the two compounds were identified by one-dimensional ((1)H, (13)C) and two-dimensional ((1)H-H and (1)H-(13)C) NMR experiments as isolimonic acid and a novel compound named as ichanexic acid. Stereochemical assignment of the protons for both the compounds was made using one-dimensional nuclear Overhauser enhancement (nOe) experiments. The identified compounds were tested for the inhibition of human colon cancer cells (HT-29) proliferation, apoptosis, and on non-cancerous (COS-1 fibroblast) cells. Cell proliferation, arrest of cell growth, and induction of apoptosis were determined by MTT assay, flow cytometry, and nuclear staining methods, respectively. The MTT assay indicated that both the compounds exhibited differential inhibition at various concentrations. Significant arrest of cell growth by isolimonoic acid was noticed within 24h of treatment on the HT-29 colon cancer cells at a concentration as low as 5.0microM (P=0.005) and by ichanexic acid at 10.0microM (P=0.011). None of the compounds exerted any apparent cytostatic effects on the non-cancerous COS-1 fibroblast cells. Both the compounds exerted nearly 4- to 5-fold increase in the counts of G2/M stage cells at 5microM indicating a potential role in the cell cycle arrest as well as possible lead structures for the development of cancer chemopreventive and therapeutic agents. To the best of our knowledge, this is the first report on isolation, identification of isolimonic acid in its native form, and compound 2 was found to a novel and identified as ichanexic acid.
Jayaprakasha
G K
GK
Vegetable & Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University, 2119 TAMU, College Station, TX 77843, USA.
Mandadi
K K
KK
Poulose
S M
SM
Jadegoud
Y
Y
Nagana Gowda
G A
GA
Patil
Bhimanagouda S
BS
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D019556:COS Cells; D049109:Cell Proliferation / Q000187:drug effects; D002522:Chlorocebus aethiops; D002957:Citrus / Q000737:chemistry*; D003110:Colonic Neoplasms / Q000188:drug therapy / Q000473:pathology; D006131:Growth Inhibitors / Q000737:chemistry / Q000494:pharmacology; D019073:HT29 Cells; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology; D012639:Seeds / Q000737:chemistry; D014315:Triterpenes / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*
D000972:Antineoplastic Agents, Phytogenic; D006131:Growth Inhibitors; D010936:Plant Extracts; D014315:Triterpenes