Identification, Synthesis, and Enzymology of Non-natural Glucosinolate Chemopreventive Candidates

• Published in: Chembiochem : a European journal of chemical biology 2008-03, Vol.9 (5), p.729-747
• Main Authors: Mays, Jared R, Weller Roska, Rachel L, Sarfaraz, Sami, Mukhtar, Hasan, Rajski, Scott R
• Format: Article
• Language: English
• Subjects: Animals; Carcinoma - drug therapy; Cell Line, Tumor; Cell Proliferation - drug effects; Chemoprevention; Chromatography, High Pressure Liquid - methods; Drug Screening Assays, Antitumor; Epithelial Cells - drug effects; glucosinolates; Glucosinolates - chemical synthesis; Glucosinolates - chemistry; Glucosinolates - pharmacology; Glycoside Hydrolases - chemistry; Humans; Hydrolysis; isothiocyanates; Isothiocyanates - chemical synthesis; Isothiocyanates - chemistry; Isothiocyanates - pharmacology; Mice; Molecular Structure; myrosinase; prodrugs; Small Molecule Libraries; Stereoisomerism; structure-activity relationships; Thiocyanates - chemical synthesis; Thiocyanates - chemistry; Thiocyanates - pharmacology; Time Factors; Water - chemistry
• ISSN: 1439-4227; 1439-7633
• DOI: 10.1002/cbic.200700586

Isothiocyanates (ITCs) are one of the many classes of breakdown products of glucosinolates found in crucifers such as broccoli and are thought to be partially responsible for the reduced risk of degenerative diseases associated with the consumption of vegetables. The production of ITCs such as L-sulforaphane is dependent on the hydrolytic bioactivities of myrosinase, localized both within vegetable tissues and within flora of the human GI tract, and is associated with important cancer chemopreventive activities. We hypothesized that novel isothiocyanates with enhanced chemopreventive properties relative to L-sulforaphane could be identified and that their glucosinolate precursors could be synthesized. From a library of 30 synthetic ITCs, we identified several with bioactivities equal or superior to those of L-sulforaphane. The corresponding non-natural glucosinolate precursors to these novel ITCs were constructed and found to be substrates for myrosinase. By utilizing a novel RP-HPLC assay to monitor myrosinase-dependent hydrolysis reactions, 2,2-diphenylethyl glucosinolate and (biphenyl-2-yl)methyl glucosinolate were shown to exhibit 26.5 and 2.8 %, respectively, of the relative activity of sinigrin and produced their corresponding ITCs in varying yields. These data support the notion that non-natural glucosinolates can act as prodrugs for novel ITCs, with a mechanism of action reliant on their hydrolytic cleavage by myrosinase. Such non-natural glucosinolates may serve as very economical chemopreventive agents for individuals at risk for cancers of and around the GI tract.