CCG-222740 is a Rho/MRTF pathway inhibitor. It decreases the activation of stellate cells in vitro and in vivo, by reducing the levels of alpha smooth muscle actin (α-SMA) expression. It also modulates inflammatory components of the pancreas in KC mice (LSL-KrasG12D/+; Pdx-1-Cre) stimulated with caerulein.
In fibroblasts, the Rho/myocardin-related transcription factor (MRTF) pathway has classically been associated with cell contractility, and CAFs (Cancer associated fibroblasts) rely on this pathway to generate tracks for cancer cell migration. This pathway is implicated in the hyper-activation of fibroblasts in fibrotic diseases and the activation of pancreatic stellate cells. CCG-222740, a small molecule, is designed as a Rho/MRTF pathway inhibitor. PSCs (Pancreatic stellate cells), isolated as previously described from the pancreas of wild type C57BL/6 mice, were cultured for 3 days to achive confluence and then treated with CCG-222740 for 6 days. Treatment with 1 μM of CCG-222740 significantly (p < 0.05) reduced the levels of α-SMA (α smooth muscle actin; a marker for stellate cell activation) and collagen 2 A levels in the PSCs. CAFs isolated from a mouse pancreatic tumor induced by a Kras mutation were treated with several concentrations of CCG-222740 for 72 hours. This treatment decreased cell viability of CAFs, with an IC50 of ~10 μM, as measured by the MTT assay. CCG-222740 also had growth inhibitory effects on human and mouse pancreatic cancer cells, inducing growth arrest at similar concentrations as the CAFs. Moreover, CCG-222740 increased the protein levels of p27 and decreased cyclin D1, while the levels of collagens I, 2a and IV, as well as α-SMA in the CAFs were decreased after treatment with CCG-222740 for 72 hours. In vivo, CCG-222740 (100 mg/kg daily) by oral gavage significantly reduced α-SMA levels in the pancreas of caerulein-stimulated KC mice (p < 0.05)[1].
CCG-222740 细胞实验
Cell Line
Concentration
Treated Time
Description
References
HEK-293T cells
30 μM
Used in affinity pulldown experiments to identify pirin as a potential target via mass spectrometry
To evaluate the effect of CCG-222740 on CAFs cell viability and collagen production, results showed that CCG-222740 decreased CAFs cell viability and reduced the levels of collagens I, 2a, IV, and α-SMA.
Sci Rep. 2019 May 8;9(1):7072
mouse pancreatic stellate cells (PSCs)
1 μM
6 days
To evaluate the effect of CCG-222740 on stellate cell activation, results showed that CCG-222740 significantly reduced the levels of α-SMA and collagen 2A.
Sci Rep. 2019 May 8;9(1):7072
M229R
10 μM
72 h
To evaluate the effect of CCG-222740 on the sensitivity of M229R cells to Vemurafenib. Results showed that CCG-222740 had a minor effect on M229R cells.
Oncogene. 2020 Feb;39(7):1466-1483.
M229P
10 μM
72 h
To evaluate the effect of CCG-222740 on the sensitivity of M229P cells to Vemurafenib. Results showed that CCG-222740 had a minor effect on M229P cells.
Oncogene. 2020 Feb;39(7):1466-1483.
UACC62R
10 μM
72 h
To evaluate the effect of CCG-222740 on the sensitivity of UACC62R cells to Vemurafenib. Results showed that CCG-222740 significantly increased the sensitivity of UACC62R cells to Vemurafenib.
Oncogene. 2020 Feb;39(7):1466-1483.
UACC62P
10 μM
72 h
To evaluate the effect of CCG-222740 on the sensitivity of UACC62P cells to Vemurafenib. Results showed that CCG-222740 increased the sensitivity of UACC62P cells to Vemurafenib.
Oncogene. 2020 Feb;39(7):1466-1483.
human conjunctival fibroblasts
1.25, 2.5, 5, 10 μM
24 h
To evaluate the effect of nanocarrier CCG-222740 on ACTA2 gene expression and cytotoxicity in human conjunctival fibroblasts. Results showed that nanocarrier CCG-222740 significantly decreased ACTA2 gene expression at 2.5, 5, and 10 μM concentrations and had no significant effect on cell viability.
J Nanobiotechnology. 2018 Nov 27;16(1):97
CCG-222740 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Mice
KC mice (LSL-KrasG12D/+; Pdx-1-Cre)
Oral gavage
100 mg/kg
Once daily for 7 days
To evaluate the effect of CCG-222740 on stellate cell activation and immune cell infiltration in KC mice, results showed that CCG-222740 significantly reduced α-SMA levels, decreased macrophage infiltration, and increased CD4 T cells and B cells.
Sci Rep. 2019 May 8;9(1):7072
New Zealand white rabbits
Glaucoma filtration surgery model
Subconjunctival injection
68 μg in 100 μl
Single administration, observed for 30 days
To evaluate the anti-fibrotic effect and safety of nanocarrier CCG-222740 in a rabbit glaucoma filtration surgery model. Results showed that nanocarrier CCG-222740 significantly prolonged bleb survival, reduced conjunctival scarring, and had no local or systemic toxicity.
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