Von Hippel-Lindau syndrome (VHL) is a component of the protein complex that includes elongin B, elongin C, and cullin-2, and possesses ubiquitin ligase E3 activity. VHL can induces hypoxia inducible factor (HIF) ubiquitination and degradation. VH-298 is a potent inhibitor of the VHL with a Kd of 90 nM as determined by isothermal titration calorimetry and 80 nM in a competitive fluorescence polarization assay. In vitro, the measured permeability of VH298 was 19.4 nm/s, and 500 μM of VH-298 did not affect the viability of HCC, Hela, RCC4-HA, RCC40-HA-VHL, U2OS and CTL cell lines, suggesting that VH298 was permeable and not toxic to cells. Treatment with 1 – 250 μM VH-298 for 2 hour induced dose-dependent accumulation of HIF-1α levels in Hela cells. In addition, VH-298 induced detectable HIF activity at concentration of 10 μM. Treatment with 100 μM VH298 elicited a HIF-dependent hypoxic response in HFF, Hela, and U2OS lines[3]. In addition, VH-298 promoted rat fibroblasts proliferation at concentration of 30 μM and 100 μM, while 10 μM and 200 μM had no significant effect on cell proliferation. Treatment with 20 μM VH-298 promoted the tubule formation of HUVECs, however, 100 μM and 200 μM of VH-298 disturbed the tubule formation of HUVECs. In vivo, administered 100 μL of 30 μM VH-298 via local injection every three days accelerated wound healing in Rats with DM[4].
作用机制
VH-298 stabilizes HIF-α and induces a hypoxic response to block VHL-HIFα interaction[3].
VH-298 细胞实验
Cell Line
Concentration
Treated Time
Description
References
U2OS cells
50 µM
16 hours
Upregulated mRNA levels of HIF-target genes CA9 and GLUT1
Nat Commun. 2016 Nov 4;7:13312
HeLa cells
10 µM
2 hours
Induced HIF-1α accumulation
Nat Commun. 2016 Nov 4;7:13312
RCC4-HA-VHL cells
150 µM
24 hours
Increased mRNA levels of EPO
Nat Commun. 2016 Nov 4;7:13312
HFF cells
100 µM
24 hours
Upregulated mRNA levels of HIF-target genes CA9 and GLUT1
Nat Commun. 2016 Nov 4;7:13312
CTL cells
100 µM
8 hours
Upregulated GLUT1 protein levels
Nat Commun. 2016 Nov 4;7:13312
Human umbilical vein endothelial cells (hUVEC)
0 µM, 10 µM, 30 µM, 100 µM, 200 µM
24 hours
To evaluate the effect of VH-298 on angiogenesis. Results showed that 30 μM VH-298 significantly promoted angiogenesis, while high doses (100 μM and 200 μM) inhibited angiogenesis.
J Diabetes Res. 2019 Feb 17;2019:1897174
Human foreskin fibroblasts (HFFs)
100 µM
24 hours
To validate the effect of VH298 on increasing VHL protein levels in HFF cells.
J Biol Chem. 2021 Aug;297(2):100910
Human foreskin fibroblasts (HFF)
100 µM
24 hours
To validate the effect of VH298 on VHL protein levels in HFF cells, results showed that VH298 treatment increased VHL protein levels.
J Biol Chem. 2021 Aug;297(2):100910
Rat fibroblasts (rFb)
0 µM, 10 µM, 30 µM, 100 µM, 200 µM
48 hours
To evaluate the effect of VH-298 on cell proliferation. Results showed that 30 μM and 100 μM VH-298 significantly promoted cell proliferation.
J Diabetes Res. 2019 Feb 17;2019:1897174
HeLa cells
50 µM
48 hours
VH298 selectively promoted peroxisome degradation without affecting mitochondria, ER, or Golgi apparatus
Molecules. 2024 Jan 18;29(2):482
HTERT RPE-1 cells
50 µM
48 hours
VH298 significantly increased the number of red-only puncta, indicating enhanced peroxisome degradation
Molecules. 2024 Jan 18;29(2):482
VH-298 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Sprague-Dawley rats
Streptozotocin (STZ)-induced hyperglycaemic rat model
Local injection
30 μM VH298
Every three days for 21 days
To evaluate the effect of VH-298 on wound healing in hyperglycaemic rats. Results showed that VH-298 significantly accelerated wound healing, increased angiogenesis and collagen deposition.
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