Hydroxyfasudil, a ROCK inhibitor, shows IC50 values of 0.73 and 0.72 μM for ROCK1 and ROCK2, respectively. It less potently inhibits PKA with an IC50 of 37 μM. Hydroxyfasudil enhances eNOS mRNA levels (EC50 of 0.8 ± 0.3 μM), and between 0-100 μM, it increases eNOS activity and stimulates NO production in human aortic endothelial cells (HAEC). At 10 μM, it prolongs the half-life of eNOS mRNA from 13 to 16 hours without affecting eNOS promoter activity [1].
体内研究
Hydroxyfasudil (10 mg/kg, i.p.) significantly increases both average and maximal voided volumes and reduces maximal detrusor pressure in SD rats [2].
Hydroxyfasudil (3 mg/kg, i.p) inhibits hypercontractility induced by norepinephrine in spontaneously hypertensive rats (SHRs), and at doses of 3 and 10 mg/kg, it significantly improves decreased penile cGMP levels in these rats [3].
体外研究
Hydroxyfasudil, a ROCK inhibitor, shows IC50 values of 0.73 and 0.72 μM for ROCK1 and ROCK2, respectively. It less potently inhibits PKA with an IC50 of 37 μM. Hydroxyfasudil enhances eNOS mRNA levels (EC50 of 0.8 ± 0.3 μM), and between 0-100 μM, it increases eNOS activity and stimulates NO production in human aortic endothelial cells (HAEC). At 10 μM, it prolongs the half-life of eNOS mRNA from 13 to 16 hours without affecting eNOS promoter activity [1].
Hydroxyfasudil/羟基法舒地尔 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Bovine aortic endothelial cells (BAEC)
0.1 to 100 μmol/L
48 hours
Hydroxyfasudil (0.1 to 100 μmol/L) did not affect eNOS promoter activity.
Stroke. 2005 Oct;36(10):2251-7
Human saphenous vein endothelial cells (HSVEC)
10 μmol/L
18 hours
Treatment with hydroxyfasudil increased eNOS mRNA expression to 156±20% (n=3, P<0.05).
Stroke. 2005 Oct;36(10):2251-7
Human umbilical vein endothelial cells (HUVEC)
10 μmol/L
18 hours
Treatment with hydroxyfasudil increased eNOS mRNA expression to 156±10% (n=3, P<0.05).
Stroke. 2005 Oct;36(10):2251-7
Human aortic endothelial cells (HAEC)
0.1 to 100 μmol/L
18 hours
Hydroxyfasudil increased eNOS mRNA and protein expression in a concentration-dependent manner, resulting in a 1.9- and 1.6-fold increase, respectively, at 10 μmol/L (P<0.05 for both). This correlated with a 1.5- and 2.3-fold increase in eNOS activity and NO production, respectively (P<0.05).
To evaluate the effect of Hydroxyfasudil on the contractile force of electrically paced guinea pig left atrium, showing no significant effect.
Br J Pharmacol. 2001 Dec;134(8):1724-30
Guinea pig right atrium
10^-7 – 10^-4 M
To evaluate the effect of Hydroxyfasudil on the spontaneous beating rate of guinea pig right atrium, showing no significant effect.
Br J Pharmacol. 2001 Dec;134(8):1724-30
Mouse motor neuron cells (NSC34)
0.3, 3, 30 nM
27 hours
To evaluate the protective effect of Hydroxyfasudil on SOD1G93A-induced motor neuron cell death. Results showed that at concentrations of 3-30 nM, Hydroxyfasudil reduced SOD1G93A-induced cell death in a concentration-dependent manner.
Br J Pharmacol. 2013 Sep;170(2):341-51
Human PKD1 cystic cell line (OX161)
1–30 μM
7 days
To test the efficacy of the ROCK inhibitor hydroxyfasudil in 3D cyst assays, results showed that hydroxyfasudil significantly reduced cyst area.
JCI Insight. 2020 Aug 20;5(16):e135385
rat basilar artery
3 μmol/L
60 mins hypoxia
ACh-induced endothelium-dependent relaxation was abolished during hypoxia and restored upon reoxygenation
J Cereb Blood Flow Metab. 2007 May;27(5):998-1009
mouse aorta
3 μmol/L
90 mins (including 60 mins hypoxia)
Partially restored endothelium-dependent ACh-induced relaxations during hypoxia, which was abolished by L-NAME, indicating eNOS mediation
J Cereb Blood Flow Metab. 2007 May;27(5):998-1009
Hydroxyfasudil/羟基法舒地尔 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Mice
Wild-type or eNOS-deficient mice
Intraperitoneal injection
1, 3, or 10 mg/kg/day（fasudil）
Once daily for 2 days
Fasudil increased cerebral blood flow to both ischemic and nonischemic brain areas, reduced cerebral infarct size by 33%, and improved neurologic deficit score by 37% (P<0.05). This correlated with inhibition of brain and vascular ROCK activity and increased eNOS expression and activity. The neuroprotective effects of fasudil were absent in eNOS-deficient mice.
Stroke. 2005 Oct;36(10):2251-7
Mice
Mesenteric ischemia-reperfusion injury model in wild-type and eNOS−/− mice
Intraperitoneal
10 mg/kg（fasudil）
Single dose 1 hour before ischemia
Fasudil decreased leukocyte recruitment and adhesion to the mesenteric endothelium after I/R injury in wild-type but not in eNOS?/? mice.
Myocardial ischemia model induced by left anterior descending coronary artery stenosis
Intravenous injection
0.1 and 0.3 mg/kg
30 minutes
To evaluate the effect of Hydroxyfasudil on ST-segment depression in a canine myocardial ischemia model, showing dose-dependent suppression of ST-segment depression and increased blood flow in the LAD-perfused endomyocardium region.
Br J Pharmacol. 2001 Dec;134(8):1724-30
Mice
SOD1G93A mutant mice (ALS model)
Administered via drinking water
30 and 100 mg/kg fasudil
From 5 weeks of age until the experimental endpoint
To evaluate the effect of Fasudil on disease progression and motor neuron loss in SOD1G93A mutant mice. Results showed that Fasudil delayed disease onset, prolonged survival time, and reduced motor neuron loss.
Br J Pharmacol. 2013 Sep;170(2):341-51
Mouse
Tetracycline-inducible kidney-specific Pkd1 mouse model (Pax8rtTA-TetO-Cre-Pkd1fl/fl)
Intraperitoneal injection
10 mg/kg/day
Once daily for 7 days
To test the inhibitory effect of hydroxyfasudil on cyst growth in vivo, results showed that treated mice had significantly reduced kidney weights and cystic indices, and increased cilia length.
JCI Insight. 2020 Aug 20;5(16):e135385
Mice (C57BL/6J and eNOS−/−)
Distal middle cerebral artery occlusion (dMCAO) model
Intraperitoneal or intravenous
10 mg/kg
Single dose, 60 mins before or 5 mins after dMCAO
In wild-type mice, hydroxyfasudil reduced the area of ischemic core and penumbra, but had no effect in eNOS?/? mice, indicating eNOS-dependent action
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