Pro-drug type gamma-Glutamylcyclotransferase (GGCT) inhibitor Pro-GA | Funakoshi

For research of glutathione metabolic cycle
Pro-GA
The cell permeable GGCT Inhibitor

GGCT (γ-Glutamylcyclotransferase, gamma-Glutamylcyclotransferase, C7orf24) is a glutathione homeostasis-related enzyme, which is highly expressed in cancer cells. While GGCT expression is low in normal cells, GGCT overexpressed cells dramatically increase its proliferation. Meanwhile, cell proliferation and migration are suppressed in GGCT KO cancer cells. This knowledge suggests that GGCT accelerates cell proliferation.
Existing GGCT inhibitors, such as N-glutaryl-L-alanine (GA), do not have cell membrane permeability, and cannot be used for cell culture and animal experiment. Pro-GA is the world’s first pro-drug type of GA, which penetrates into cells with inhibition activity to GGCT.

This product has been commercialized with the support of Kyoto Pharmaceutical University.

Fig.1 GA-release efficiency of Pro-GA in cells
MCF7 cells were treated with 100 μM of fluorescent NBD-conjugated Pro-GA for 0-60 min. After washing, cell lysates were prepared and analyzed by HPLC to evaluate GA amount released from Pro-GA. NBD-conjugated Pro-GA was immediately incorporated into cells within a few minutes and converted to GA in the cells.

Fig.2 Inhibition of proliferation in cancer cells
Cells were cultured in serum starve condition with DMSO (as control), GA or Pro-GA for the indicated days. Cell proliferation was measured by WST cell viability assay. Pro-GA specifically suppressed proliferation of cancer cells.

Fig.3 Stability of Pro-GA in culture medium at room temperature
Stabilities of fluorescent NBD-conjugated Pro-GA in PBS or 10% FBS/DMEM were tested by HPLC analysis. While conversion of Pro-GA to GA was slowly in PBS (t1/2>5 hours), Pro-GA was quickly converted into GA in 10% FBS/DMEM by esterase activity in FBS (t1/2~50 min).
Please prepare Pro-GA solution in assay medium just before use.

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