Isolation, purification and characterization of naturally derived Crocetin beta-d-glucosyl ester from Crocus sativus L. against breast cancer and its binding chemistry with ER-alpha/HDAC2

Mudasir A Mir; Shabir Ahmad Ganai; Sheikh Mansoor; Sumira Jan; P Mani; Khalid Z Masoodi; Henna Amin; Muneeb U Rehman; Parvaiz Ahmad

doi:10.1016/j.sjbs.2020.01.018

Isolation, purification and characterization of naturally derived Crocetin beta-d-glucosyl ester from Crocus sativus L. against breast cancer and its binding chemistry with ER-alpha/HDAC2

Saudi J Biol Sci. 2020 Mar;27(3):975-984. doi: 10.1016/j.sjbs.2020.01.018. Epub 2020 Jan 27.

Authors

Mudasir A Mir¹, Shabir Ahmad Ganai², Sheikh Mansoor³, Sumira Jan⁴, P Mani⁵, Khalid Z Masoodi¹, Henna Amin⁶, Muneeb U Rehman⁷, Parvaiz Ahmad⁸

Affiliations

¹ Division of Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar 190025, Jammu and Kashmir, India.
² Division of Basic Sciences and Humanities, Faculty of Agriculture, SKUAST-Kashmir, Wadura, Sopore 193201, Jammu & Kashmir, India.
³ Division of Biochemistry, Sher-e-Kashmir University of Agricultural Sciences and Technology, Jammu 180009, J&K, India.
⁴ Division of Basic Science and Humanities, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar 190025, Jammu and Kashmir, India.
⁵ Department of Biotechnology, Annai College of Arts & Science, Kumbakonam, Tamil Nadu 612503, India.
⁶ Department of Pharmaceutical Sciences, Faculty of Applied Science and Technology, University of Kashmir, Srinagar, J&K 190006, India.
⁷ Department of Clinical Pharmacy, College of Pharmacy, King Saudi University, P. O. Box. 2460, Riyadh 11451, Saudi Arabia.
⁸ Botany and Microbiology Department, College of Science, King Saudi University, P. O. Box. 2460, Riyadh 11451, Saudi Arabia.

Abstract

Saffron plant (Crocus sativus L.) is being used as a source of saffron spice and medicine to cure or prevent different types of diseases including cancers. We report the isolation, characterization of bioactive small molecule ([crocetin (β-d-glucosyl) ester] from the leaf biowastes of saffron plant of Kashmir, India. MTTC assay and Bio-autography aided approach were used to assess anti-oxidant activity and anti-cancer properties of crocin (s) against DPPH free radical and breast cancer cell line respectively. Crocetin beta-d-glucosyl ester restrained proliferation of human breast adeno-carcinoma cell model (MCF-7) without significantly affecting normal cell line (L-6). Further studies involving molecular mechanics generalized born surface area and molecular docking showed that crocetin beta-d-glucosyl ester exhibits strong affinity for estrogen receptor alpha and histone deacetylase 2 (crucial receptors involved in breast cancer signalling) as evidenced by the negative docking score and binding free energy (BFE) values. Therefore, crocetin beta-d-glucosyl ester from Crocus sativus biowastes showed antiproliferative effect possibly by inhibiting estrogen receptor alpha and HDAC2 mediated signalling cascade.

Keywords: Antioxidant; Breast cancer; Crocetin beta-d-glucosyl ester; DMEM, Dulbecco’s Modified Eagle’s Medium; DPPH, 2,2-diphenyl-1-picrylhydrazyl; FBS, Fetal Bovine serum; FTIR, Fourier-transform infrared spectroscopy; Floral biowastes; LC-MS/MS, Liquid chromatography–mass spectrometry; MTT, 3-(4,5–dimethyl thiazol–2–yl)–5–diphenyltetrazolium bromide; Molecular docking; NMR, Nuclear magneticresonance; Saffron; TLC, Thin layer chromatography; UV, Ultra violet.