Supplementary information
Stability, Permeability and Growth-Inhibitory Properties of
Gonadotropin-Releasing Hormone Liposaccharides
Daryn Goodwin1, Pegah Varamini1, Pavla Simerska1, Istvan Toth1,2*
1
School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia
4072, Queensland, Australia; 2The School of Pharmacy, Pharmacy Australia Centre of
Excellence, Wooloongabba 4102, Queensland, Australia
*Corresponding author:
School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia,
Queensland, Australia, 4072
Tel: +61 7 33469892
Fax: +61 7 33654273
email: i.toth@uq.edu.au
Figure S1. Lipidation strategy using activated lipoamino acid-NHS ester on Tyr.
Figure S2. Stability of C8 conjugated GnRH(glyco)lipids in Caco-2 cell homogenates (mean
± SEM). Addition of lipoamino acids and carbohydrates conferred increases in half-life
during incubation. Peptides modified with dimeric addition of lipoamino acid were detected
in the highest concentrations at completion of the assay.
Figure S3. The generation of native GnRH (10) in the receiver chamber demonstrated
indicated terminal cyclisation of [E]GnRH (1) during incubation with Caco-2 cell monolayers
(mean ± SD).
Figure S4. The trans-epithelial electrical resistance (TEER) of the cell monolayer
measured after incubation with (glyco)lipopeptides (mean±SD). TEER was measured at t0
(before addition of conjugates), at 3 h (after removal of conjugates), and 16 h (overnight).
Figure S5. Viability of PC-3 cells as a percentage of cells treated with PBS (mean±SEM)
after 48h incubation with C8- and C12-(glyco)lipopeptides (at 100 µg/mL) in 10% DMSO.
Statistical analysis was performed using a one-way ANOVA followed by the Dunnett’s post
hoc test and compared to DMSO group (*, p > 0.05; **, p < 0.01; ***, p < 0.001).
a
b
Figure S6. Viability of SKOV-3 cells as a percentage of cells treated with PBS (mean±SEM)
after 48h incubation with C8- (a) and C12-(glyco)lipopeptides (b) in 10% DMSO. Statistical
analysis was performed using a one-way ANOVA followed by the Dunnett’s post hoc test
and compared to DMSO group (*, p > 0.05;***, p<0.001).