FJ9 is a NHERF1/PDZ inhibitor with human NHERF1 PDZ1 IC50 1540 μM, NHERF1 PDZ2 IC50 160 μM, and Frizzled-7-Dishevelled PDZ complex K i 10 μM. FJ9 binds ligand-binding pockets of NHERF1 PDZ domains to block cognate ligand interactions, disrupts Frizzled-7-Dishevelled interactions, and down-regulates canonical Wnt signaling. FJ9 induces apoptosis in cancer cells with intact β-catenin signaling. FJ9 can be used for the research of non-small cell lung cancer, melanoma [1] [2].
In Vivo, FJ9 (50 mg/kg; i.p.; daily; 14 days) significantly inhibits H460 non-small cell lung cancer xenograft growth in nude mice without causing significant weight loss[2].
In Vitro, FJ9 (Compound 1) inhibits the interaction between GST-fused human NHERF1 PDZ1 domain and biotinylated β2AR carboxy-terminal peptide with an IC50 of 1540 μM[1]. FJ9 inhibits the interaction between GST-fused human NHERF1 PDZ2 domain and biotinylated CFTR carboxy-terminal peptide with an IC50 of 160 μM[1]. FJ9 (0.1-1000 μM) competitively inhibits Frz7 peptide binding to purified human DVL1 PDZ domain with a Ki of 63 μM and to purified human DVL3 PDZ domain with a Ki of 29 μM[2]. FJ9 (100-300 μM; 24 hours) penetrates HEK293T cells and disrupts the intracellular interaction between HA-tagged mouse DVL1 and myc-tagged human Frz7 at concentrations of 100 μM and 300 μM[2]. FJ9 (100-300 μM; 48 hours) dose-dependently down-regulates cytosolic β-catenin levels in hTERT-immortalized LP9 mesothelial cells stably expressing full-length mouse DVL1, but not in control LP9 cells or LP9 cells expressing PDZ domain-deleted mouse DVL1[2]. FJ9 (100-300 μM; 24 hours) dose-dependently inhibits constitutive Tcf transcriptional activity in HCT116 p53-/- colon cancer cells, reducing TOPflash reporter activity while minimally affecting FOPflash reporter activity[2]. FJ9 (30-100 μM; 1-2 days) down-regulates canonical Wnt signaling target proteins (cyclin D1, c-myc, survivin) and mRNA (cyclin D1, survivin) in HCT116 p53-/- colon cancer cells, and down-regulates survivin protein in NCI-H1703 non-small cell lung cancer cells[2]. FJ9 (30-100 μM; 24 hours) does not affect JNK activation in mDVL1-transfected HeLa cells after 24 hours of treatment at 30 μM and 100 μM[2]. FJ9 (86-100 μM; 3 to 6 days) induces significant apoptosis in LOX melanoma, NCI-H460 non-small cell lung cancer, and NCI-H1703 non-small cell lung cancer cells, but has no apoptotic effect on NCI-H28 mesothelioma cells or normal NHBE and SAEC cells, after 3 to 6 days of treatment at 86 to 100 μM[2].
References:
[1]. Mayasundari A, et al. Rational design of the first small-molecule antagonists of NHERF1/EBP50 PDZ domains. Bioorg Med Chem Lett. 2008;18(3):942-945.
[2]. Fujii N, et al. An antagonist of dishevelled protein-protein interaction suppresses beta-catenin-dependent tumor cell growth. Cancer Res. 2007;67(2):573-579.
In Vivo, FJ9 (50 mg/kg; i.p.; daily; 14 days) significantly inhibits H460 non-small cell lung cancer xenograft growth in nude mice without causing significant weight loss[2].
In Vitro, FJ9 (Compound 1) inhibits the interaction between GST-fused human NHERF1 PDZ1 domain and biotinylated β2AR carboxy-terminal peptide with an IC50 of 1540 μM[1]. FJ9 inhibits the interaction between GST-fused human NHERF1 PDZ2 domain and biotinylated CFTR carboxy-terminal peptide with an IC50 of 160 μM[1]. FJ9 (0.1-1000 μM) competitively inhibits Frz7 peptide binding to purified human DVL1 PDZ domain with a Ki of 63 μM and to purified human DVL3 PDZ domain with a Ki of 29 μM[2]. FJ9 (100-300 μM; 24 hours) penetrates HEK293T cells and disrupts the intracellular interaction between HA-tagged mouse DVL1 and myc-tagged human Frz7 at concentrations of 100 μM and 300 μM[2]. FJ9 (100-300 μM; 48 hours) dose-dependently down-regulates cytosolic β-catenin levels in hTERT-immortalized LP9 mesothelial cells stably expressing full-length mouse DVL1, but not in control LP9 cells or LP9 cells expressing PDZ domain-deleted mouse DVL1[2]. FJ9 (100-300 μM; 24 hours) dose-dependently inhibits constitutive Tcf transcriptional activity in HCT116 p53-/- colon cancer cells, reducing TOPflash reporter activity while minimally affecting FOPflash reporter activity[2]. FJ9 (30-100 μM; 1-2 days) down-regulates canonical Wnt signaling target proteins (cyclin D1, c-myc, survivin) and mRNA (cyclin D1, survivin) in HCT116 p53-/- colon cancer cells, and down-regulates survivin protein in NCI-H1703 non-small cell lung cancer cells[2]. FJ9 (30-100 μM; 24 hours) does not affect JNK activation in mDVL1-transfected HeLa cells after 24 hours of treatment at 30 μM and 100 μM[2]. FJ9 (86-100 μM; 3 to 6 days) induces significant apoptosis in LOX melanoma, NCI-H460 non-small cell lung cancer, and NCI-H1703 non-small cell lung cancer cells, but has no apoptotic effect on NCI-H28 mesothelioma cells or normal NHBE and SAEC cells, after 3 to 6 days of treatment at 86 to 100 μM[2].
References:
[1]. Mayasundari A, et al. Rational design of the first small-molecule antagonists of NHERF1/EBP50 PDZ domains. Bioorg Med Chem Lett. 2008;18(3):942-945.
[2]. Fujii N, et al. An antagonist of dishevelled protein-protein interaction suppresses beta-catenin-dependent tumor cell growth. Cancer Res. 2007;67(2):573-579.