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Small molecule proprotein convertase inhibitors for inhibition of embryo implantation.

Ho H, Singh H, Heng S, Nero TL, Paule S, Parker MW, Johnson AT, Jiao GS, Nie G - PLoS ONE (2013)

Bottom Line: Compound 1o is unique as it appears the most lipophilic among the five compounds.Our findings also highlight that human cell-based functional models are vital to complement the biochemical and in silico analyses in the selection of promising drug candidates.Further investigations for compound 1o are warranted in animal models to test its utility as an implantation-inhibiting contraceptive drug.

View Article: PubMed Central - PubMed

Affiliation: Prince Henry's Institute of Medical Research, Clayton, Victoria, Australia ; Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.

ABSTRACT
Uterine proprotein convertase (PC) 6 plays a critical role in embryo implantation and is pivotal for pregnancy establishment. Inhibition of PC6 may provide a novel approach for the development of non-hormonal and female-controlled contraceptives. We investigated a class of five synthetic non-peptidic small molecule compounds that were previously reported as potent inhibitors of furin, another PC member. We examined (i) the potency of these compounds in inhibiting PC6 activity in vitro; (ii) their binding modes in the PC6 active site in silico; (iii) their efficacy in inhibiting PC6-dependent cellular processes essential for embryo implantation using human cell-based models. All five compounds showed potent inhibition of PC6 activity in vitro, and in silico docking demonstrated that these inhibitors could adopt a similar binding mode in the PC6 active site. However, when these compounds were tested for their inhibition of decidualization of primary human endometrial stromal cells, a PC6-dependent cellular process critical for embryo implantation, only one (compound 1o) showed potent inhibition. The lack of activity in the cell-based assay may reflect the inability of the compounds to penetrate the cell membrane. Because compound's lipophilicity is linked to cell penetration, a measurement of lipophilicity (logP) was calculated for each compound. Compound 1o is unique as it appears the most lipophilic among the five compounds. Compound 1o also inhibited another crucial PC6-dependent process, the attachment of human trophoblast spheroids to endometrial epithelial cells (a model for human embryo attachment). We thus identified compound 1o as a potent small molecule PC6 inhibitor with pharmaceutical potential to inhibit embryo implantation. Our findings also highlight that human cell-based functional models are vital to complement the biochemical and in silico analyses in the selection of promising drug candidates. Further investigations for compound 1o are warranted in animal models to test its utility as an implantation-inhibiting contraceptive drug.

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Inhibition of PC6 activity by small molecule compounds.The data are expressed as percent reduction of PC6 activity based on the rate of substrate hydrolysis relative to the control. Each value represents mean ± SEM of at least two independent experiments.
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pone-0081380-g002: Inhibition of PC6 activity by small molecule compounds.The data are expressed as percent reduction of PC6 activity based on the rate of substrate hydrolysis relative to the control. Each value represents mean ± SEM of at least two independent experiments.

Mentions: Of the five compounds, four of them (1e, 1f, 1g and 1n) were previously shown to be potent inhibitors of both human furin and PC6 in vitro[15] but compound 1o had not been screened against PC6. We confirmed that all five compounds inhibit in vitro rhPC6 hydrolysis of the fluorogenic peptide substrate pERTKR-AMC [19]. At 10 µM, compounds 1e, 1f, 1g and 1n inhibited rhPC6 ≥ 90%, whereas compound 1o had a slightly lower inhibitory potency of 85% (Figure 2).


Small molecule proprotein convertase inhibitors for inhibition of embryo implantation.

Ho H, Singh H, Heng S, Nero TL, Paule S, Parker MW, Johnson AT, Jiao GS, Nie G - PLoS ONE (2013)

Inhibition of PC6 activity by small molecule compounds.The data are expressed as percent reduction of PC6 activity based on the rate of substrate hydrolysis relative to the control. Each value represents mean ± SEM of at least two independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3852413&req=5

pone-0081380-g002: Inhibition of PC6 activity by small molecule compounds.The data are expressed as percent reduction of PC6 activity based on the rate of substrate hydrolysis relative to the control. Each value represents mean ± SEM of at least two independent experiments.
Mentions: Of the five compounds, four of them (1e, 1f, 1g and 1n) were previously shown to be potent inhibitors of both human furin and PC6 in vitro[15] but compound 1o had not been screened against PC6. We confirmed that all five compounds inhibit in vitro rhPC6 hydrolysis of the fluorogenic peptide substrate pERTKR-AMC [19]. At 10 µM, compounds 1e, 1f, 1g and 1n inhibited rhPC6 ≥ 90%, whereas compound 1o had a slightly lower inhibitory potency of 85% (Figure 2).

Bottom Line: Compound 1o is unique as it appears the most lipophilic among the five compounds.Our findings also highlight that human cell-based functional models are vital to complement the biochemical and in silico analyses in the selection of promising drug candidates.Further investigations for compound 1o are warranted in animal models to test its utility as an implantation-inhibiting contraceptive drug.

View Article: PubMed Central - PubMed

Affiliation: Prince Henry's Institute of Medical Research, Clayton, Victoria, Australia ; Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.

ABSTRACT
Uterine proprotein convertase (PC) 6 plays a critical role in embryo implantation and is pivotal for pregnancy establishment. Inhibition of PC6 may provide a novel approach for the development of non-hormonal and female-controlled contraceptives. We investigated a class of five synthetic non-peptidic small molecule compounds that were previously reported as potent inhibitors of furin, another PC member. We examined (i) the potency of these compounds in inhibiting PC6 activity in vitro; (ii) their binding modes in the PC6 active site in silico; (iii) their efficacy in inhibiting PC6-dependent cellular processes essential for embryo implantation using human cell-based models. All five compounds showed potent inhibition of PC6 activity in vitro, and in silico docking demonstrated that these inhibitors could adopt a similar binding mode in the PC6 active site. However, when these compounds were tested for their inhibition of decidualization of primary human endometrial stromal cells, a PC6-dependent cellular process critical for embryo implantation, only one (compound 1o) showed potent inhibition. The lack of activity in the cell-based assay may reflect the inability of the compounds to penetrate the cell membrane. Because compound's lipophilicity is linked to cell penetration, a measurement of lipophilicity (logP) was calculated for each compound. Compound 1o is unique as it appears the most lipophilic among the five compounds. Compound 1o also inhibited another crucial PC6-dependent process, the attachment of human trophoblast spheroids to endometrial epithelial cells (a model for human embryo attachment). We thus identified compound 1o as a potent small molecule PC6 inhibitor with pharmaceutical potential to inhibit embryo implantation. Our findings also highlight that human cell-based functional models are vital to complement the biochemical and in silico analyses in the selection of promising drug candidates. Further investigations for compound 1o are warranted in animal models to test its utility as an implantation-inhibiting contraceptive drug.

Show MeSH
Related in: MedlinePlus