Protein kinase D (PKD, also known as PKCmu) is closely related to the protein kinase C superfamily but is differentially regulated and has a distinct catalytic domain that shares homology with Ca(2+)-dependent protein kinases. PKD is highly expressed in hematopoietic cells and undergoes rapid and sustained activation upon stimulation of immune receptors. PKD is regulated through phosphorylation by protein kinase C (PKC)[1]. CRT0066101 dihydrochloride is a potent and specific PKD inhibitor with IC50 values of 1, 2.5 and 2 nM for PKD1, 2, and 3 respectively. CRT0066101 specifically blocks PKD1/2 activity and does not suppress PKCα/PKCβ/PKCε activity in multiple cancer cell types including A549 (lung) and MiaPaCa-2 (pancreas). CRT0066101 significantly inhibits Panc-1 cell proliferation with an IC50 value of 1 µM. Treatment with CRT0066101 results in a 6-10 fold induction of apoptosis in Panc-1 cells. CRT0066101 significantly reduces cell proliferation of Colo357, Panc-1, MiaPaCa-2, and AsPC-1 cells but has a modest effect in Capan-2 cells. CRT0066101 (5 µM) blocks both the basal and NT-induced pS916-PKD1/2 (activated PKD1/2) in Panc-1 and Panc-28 cells. CRT0066101 reduces PKD-dependent NF-κB activation and NF-κB-dependent gene expressions in Panc-1. Optimal therapeutic concentrations (8 µM) of CRT0066101 are detectable 6 h after oral administration of this drug. CRT0066101 given orally (80 mg/kg/day) for 28 days significantly abrogates PaCa growth in Panc-1 subcutaneous xenograft model. Activated PKD1/2 expression in the treated tumor-explants is significantly inhibited with peak tumor concentration (12 µM) of CRT0066101 achieved within 2 h after oral administration. Further, CRT0066101 given orally (80 mg/kg/day) for 21 days in Panc-1 orthotopic model potently blocks tumor growth in vivo. CRT0066101 significantly reduces Ki-67+ proliferation index, increases TUNEL+ apoptotic cells (p<0.05), and abrogates expression of NF-κB-dependent proteins including cyclin D1, survivin, and cIAP-1[2].
CRT0066101 2HCl 细胞实验
Cell Line
Concentration
Treated Time
Description
References
PC3-ML cells
5 µM
22 h
Inhibited invasion of PC3-ML prostate tumor cells
Am J Pathol. 2023 May;193(5):624-637
PC3-ML cells
5 µM
24 h
Inhibited migration of PC3-ML prostate tumor cells
Am J Pathol. 2023 May;193(5):624-637
MDA-MB-231
1 µM
12 h
Inhibits PRKD3 kinase activity, reduces CLU protein levels
Adv Sci (Weinh). 2021 Jan 6;8(4):2003205
MDA-MB-468
1 µM
12 h
Inhibits PRKD3 kinase activity, reduces CLU protein levels
Adv Sci (Weinh). 2021 Jan 6;8(4):2003205
C4-2B cells
5 µM
24 h
Inhibited migration of C4-2B cells
Am J Pathol. 2023 May;193(5):624-637
Porcine primary chondrocytes
10 µM
24 h
To evaluate the protective effects of CRT0066101 against IL-1β-induced apoptosis and inflammation in chondrocytes. Results showed that CRT0066101 significantly reduced IL-1β-induced cytotoxicity, apoptosis, and inflammation, and restored normal morphology and viability of chondrocytes.
Int J Nanomedicine. 2019 Nov 11;14:8835-8846
Human uroepithelial cell line SV-HUC
5 µM
6 days
CRT0066101 had minimal effect on the viability of SV-HUC cells, with a viability rate of 92.13%.
Cell Mol Life Sci. 2018 Mar;75(5):939-963
Bladder cancer cell line UMUC1
0.625-20 µM
4 days
CRT0066101 inhibited the proliferation of UMUC1 cells with an IC50 value of 0.4796 µM.
Cell Mol Life Sci. 2018 Mar;75(5):939-963
Bladder cancer cell line TCCSUP
0.625-20 µM
4 days
CRT0066101 inhibited the proliferation of TCCSUP cells with an IC50 value of 1.4300 µM.
Cell Mol Life Sci. 2018 Mar;75(5):939-963
Bladder cancer cell line T24T
0.625-20 µM
4 days
CRT0066101 inhibited the proliferation of T24T cells with an IC50 value of 0.3333 µM.
Cell Mol Life Sci. 2018 Mar;75(5):939-963
Bladder cancer cell line T24
0.625-20 µM
4 days
CRT0066101 inhibited the proliferation of T24 cells with an IC50 value of 0.4782 µM.
Cell Mol Life Sci. 2018 Mar;75(5):939-963
HCC1954
2.5 μM
16 h
CRT0066101 significantly reduced the migration and invasion capabilities of HCC1954 cells.
Mol Cancer Ther. 2015 Jun;14(6):1306-16
MDA-MB-231
2.5 μM
60 h
CRT0066101 significantly reduced the proliferation of MDA-MB-231 cells.
Mol Cancer Ther. 2015 Jun;14(6):1306-16
Bone marrow-derived macrophages
2 μM
2 h
To investigate the effect of CRT0066101 on LPS-induced TGF-β1 expression, results showed that CRT significantly inhibited TGF-β1 expression.
Lab Invest. 2023 Feb;103(2):100018
MiaPaCa-2 cells
2.5 µM
1 hour
CRT0066101 inhibited PKD activation in MiaPaCa-2 cells and prevented the increase in CTGF, CYR61, and CXCL5 mRNA levels induced by insulin and neurotensin.
Mol Cancer Res. 2017 Jul;15(7):929-941
PANC-1 cells
2.5 µM
1 hour
CRT0066101 inhibited PKD activation in PANC-1 cells and prevented the increase in CTGF, CYR61, and CXCL5 mRNA levels induced by insulin and neurotensin.
Mol Cancer Res. 2017 Jul;15(7):929-941
THP-1-derived macrophages
2.5 µM
24 h
To evaluate the inhibitory effect of CRT0066101 on LPS-induced inflammatory cytokine production. Results showed that CRT0066101 significantly reduced the protein and mRNA levels of LPS-induced cytokines (e.g., IL-6, TNF-α, and IL-1β).
Int Immunopharmacol. 2023 Jul;120:110240
GM133
0.5 and 1 μM
3 days
Inhibited cell proliferation, with inhibition rates between 45% and 81%
Neuro Oncol. 2014 Jul;16(7):933-45
U87MG
0.5 and 1 μM
3 days
Inhibited cell proliferation, with inhibition rates between 45% and 81%
Neuro Oncol. 2014 Jul;16(7):933-45
CRT0066101 2HCl 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Nude mice
TNBC cell line xenograft murine models
Subcutaneous injection
50 mg/kg
Weekly for several weeks
Significantly suppresses TNBC tumor growth
Adv Sci (Weinh). 2021 Jan 6;8(4):2003205
Athymic NCr-nu/nu mice
UMUC1 subcutaneous xenograft model
Oral gavage
120 mg/kg
3 times per week for 25 days
CRT0066101 significantly inhibited the growth of UMUC1 xenografts, with tumor volumes of 835.83 mm3 in the treated group compared to 1943.08 mm3 in the control group at day 25.
Cell Mol Life Sci. 2018 Mar;75(5):939-963
NOD scid mice
Orthotopic mammary fat pad tumor model
Oral
80 mg/kg
Every other day for eight weeks
CRT0066101 significantly reduced primary tumor growth and metastasis.
Mol Cancer Ther. 2015 Jun;14(6):1306-16
Nude mice
HCT116 xenograft model
Oral
40, 80, 120 mg/kg
Once daily for 3 weeks
Significantly inhibited tumor growth, with tumor volume reductions of 55.6%, 65.2%, and 69.5%
Mol Cancer Ther. 2014 May;13(5):1130-41
C57BL/6 mice
Bleomycin-induced dermal fibrosis model
Oral
80 mg/kg
Once daily for 4 weeks
To evaluate the therapeutic effect of CRT0066101 on bleomycin-induced dermal fibrosis, results showed that CRT significantly reduced dermal thickness, α-SMA, collagen, and IL-6 expression.
Lab Invest. 2023 Feb;103(2):100018
C57BL/6 mice
LPS-induced acute lung injury model
Intraperitoneal injection
10 mg/kg body weight
Once every other day for a total of three treatments
To evaluate the protective effect of CRT0066101 on LPS-induced acute lung injury. Results showed that CRT0066101 significantly reduced LPS-induced alveolar wall swelling and inflammatory cell infiltration, decreased the number of inflammatory cells in BALF, and lowered the levels of inflammatory cytokines (IL-1β, TNF-α, and IL-6).
Int Immunopharmacol. 2023 Jul;120:110240
Nude mice
PC3-ML cell bone metastasis model
Oral gavage
80 mg/kg
Once daily for 4 weeks
Inhibited metastasis of prostate cancer cells to bone
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