The DNA methyltransferase (DNMT) family are enzymes that can mediate the DNA methylation. Inhibiting the DNMTs have shown particular promise in reducing the formation of tumors[6]. Zebularine is a second-generation inhibitor of DNMT with IC50 value of 95 nM on DNMT1[7]. The TFK-1 and HuCCT1 cells were treated with zebularine from 100 - 1000 µM for 72 hours. The cell viability was significantly reduced in a dose dependent manner measured by WST assay. The protein expression level of DNMT1, DNMT3a, DNMT3b examined by western blotting assay was significantly inhibited for both cell lines at concentration of 400 µM and above[8]. BALB/c nude mice with tumor were treated with 10, 50 and 100 mg/kg zebularine via oral gavage in a solution of 0.45% saline every four days for 20 days. The tumor growth was reduced in a dose dependent manner and the cell death in the tumor mass via apoptosis was induced by the treatement of zebularine as measured by the TUNEL assay[9].
作用机制
Zebularine could form tight covalent complexes between the DNMT proteins and zebularine-substitute DNA[10].
Zebularine/泽布拉林 细胞实验
Cell Line
Concentration
Treated Time
Description
References
HaCaT cells
1 μg/mL
48 h
To investigate the effect of Zebularine on the proliferation and cytotoxicity of HaCaT cells. Results showed that Zebularine slightly stimulated proliferation at low concentrations (0.1 and 1.0 μg/mL) but inhibited proliferation at higher concentrations.
EBioMedicine. 2019 Aug;46:317-329.
46BR.1N cells
1 μg/mL
48 h
To investigate the effect of Zebularine on the proliferation and cytotoxicity of 46BR.1N cells. Results showed that Zebularine slightly stimulated proliferation at low concentrations (0.1 and 1.0 μg/mL) but inhibited proliferation at higher concentrations.
EBioMedicine. 2019 Aug;46:317-329.
Arabidopsis thaliana cells
20 µM
10 days
Screening for mutants sensitive to Zebularine, revealing that SMC6B mutants are highly sensitive to Zebularine
Plant Cell. 2023 Apr 20;35(5):1532-1547.
Diabetic LEC
1–20 µM
24 h
Zebularine treatment significantly increased Wnt-5a levels and stimulated wound healing in a dose-dependent manner, with a 1.6-fold increase by 24 h.
Diabetologia. 2023 Oct;66(10):1943-1958.
HL60 cells
50 or 100 µM
96 h
To investigate the effect of Zebularine on HL60 cell differentiation, results showed that Zebularine significantly increased the expression of the CD11b differentiation marker.
Cells. 2020 Aug 29;9(9):1991.
Zebularine/泽布拉林 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Mice
Unilateral ureteral obstruction model
Intraperitoneal injection
225 mg/kg
Once daily for 3 or 7 days
Zebularine significantly attenuated renal tubulointerstitial fibrosis and inflammation induced by unilateral ureteral obstruction.
Int J Mol Sci. 2022 Nov 14;23(22):14045
BALB/c mice
Oral cancer model
Oral
1 mg/ml
Daily, starting from PID 4
To evaluate the analgesic effect of Zebularine in an oral cancer model, results showed that Zebularine significantly reduced tumor growth and produced mechanical and thermal antinociception.
Clin Cancer Res. 2014 Sep 15;20(18):4882-4893
Mice
Ear pinna injury model
Intraperitoneal injection
1000 mg/kg
7 injections over 10 days
To investigate the effect of Zebularine on ear pinna wound regeneration in mice. Results showed that Zebularine significantly promoted ear hole repair, with the mean hole area decreasing by 83.2 ± 9.4% in treated mice compared to 43.6 ± 15.4% in control mice.
EBioMedicine. 2019 Aug;46:317-329.
Mice
MMTV-PyMT transgenic mice
Drinking water
5 mg/mL
Daily, for 48 days
Zebularine significantly delayed mammary tumor growth, and after 48 days of treatment, the tumors were predominantly necrotic, with a high apoptotic index observed by TUNEL assay as early as 13 days following treatment.
Mol Cancer Ther. 2012 Feb;11(2):370-82
Mice
Experimental autoimmune uveitis (EAU) model
Intraperitoneal injection
10 µg/g
Once daily for 7 consecutive days
To evaluate the effect of zebularine on intraocular inflammation in EAU mice, results showed that zebularine significantly alleviated intraocular inflammation and retinal tissue damage.
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