Ki: 0.33 μM (ROCK1)[1] IC50: 0.158 μM (ROCK2), 4.58 μM (PKA), 12.30 μM (PKC), 1.650 μM (PKG)[1]

Rho kinase (ROCK) plays a critical role in actin cytoskeleton organization and is involved in diverse fundamental cellular functions such as contraction and gene expression. Fasudil, a ROCK inhibitor, has been clinically applied since 1995 for the treatment of subarachnoid hemorrhage (SAH) in Japan. Increasing evidences indicate that fasudil could exhibit markedly therapeutic effect on central nervous system (CNS) disorders, such as Alzheimer's disease. Areas covered: This article summarizes results from supporting evidence for the potential therapy for fasudil against a variety of CNS diseases. And the properties of its analogs are also summarized. Expert opinion: Current therapies against CNS disorders are only able to attenuate the symptoms and fail in delaying or preventing disease progression and new approaches with disease-modifying activity are desperately needed. The dramatic effects of fasudil in animal models and/or clinical applications of CNS disorders make it a promising strategy to overcome CNS disorders in human beings. Given the complex pathology of CNS disorders, further efforts are necessary to develop multifunctional fasudil derivatives or combination strategies with other drugs in order to exert more powerful effects with minimized adverse effects in the combat of CNS disorders[1].

Dysfunction of the blood-brain barrier (BBB) and blood-spinal cord barrier (BSCB) is a primary characteristic of multiple sclerosis (MS). We evaluated the protective effects of fasudil, a selective ROCK inhibitor, in a model of experimental autoimmune encephalomyelitis (EAE) that was induced by guinea-pig spinal cord. In addition, we studied the effects of fasudil on BBB and BSCB permeability. We found that fasudil partly alleviated EAE-dependent damage by decreasing BBB and BSCB permeability. These results provide rationale for the development of selective inhibitors of Rho kinase as a novel therapy for MS. https://pubmed.ncbi.nlm.nih.gov/21978848/

Oral LD50 in rats: 335 mg/kg. Sensory organs and special senses: ptosis; Behavior: tremor; Behavior: seizures or effect on the epilepsy threshold. Yakuri to Chiryo. Pharmacology and Therapeutics, 20 (Supplement). Subcutaneous LD50 in rats: 123 mg/kg. Sensory organs and special senses: ptosis; Behavior: tremor; Behavior: seizures or effect on the epilepsy threshold. Pharmacology and Therapeutics, 20 (Supplement). Intravenous LD50 in rats: 59900 ug/kg. Sensory organs and special senses: ptosis; Behavior: seizures or effect on the epilepsy threshold; Gastrointestinal tract: changes in salivary gland structure or function. Pharmacology and Therapeutics, 20(Supplement)
Oral LD50 in mice: 274 mg/kg. Sensory organs and special senses: ptosis; Behavior: altered sleep duration (including altered righting reflex); Behavior: seizures or effect on epilepsy threshold. Yakuri to Chiryo. Pharmacology and Therapeutics., 20(Supplement)
Subcutaneous LD50 in mice: 124 mg/kg. Sensory organs and special senses: ptosis; Behavior: altered sleep duration (including altered righting reflex); Behavior: seizures or effect on epilepsy threshold. Yakuri to Chiryo. Pharmacology and Therapeutics, 20(Supplement)

[1].Fasudil and its analogs: a new powerful weapon in the long war against central nervous system disorders? Expert Opin Investig Drugs. 2013 Apr;22(4):537-50.

[2].The effects of fasudil on the permeability of the rat blood-brain barrier and blood-spinal cordbarrier following experimental autoimmune encephalomyelitis. J Neuroimmunol. 2011 Oct 28;239(1-2):61-7.

[3].Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature. 1997 Oct 30;389(6654):990-4.

[4].Fasudil hydrochloride hydrate, a Rho-kinase (ROCK) inhibitor, suppresses collagen production and enhances collagenase activity in hepatic stellate cells. Liver Int. 2005 Aug;25(4):829-38.

[5].Inhibition of the activity of Rho-kinase reduces cardiomyocyte apoptosis in heart ischemia/reperfusion via suppressing JNK-mediated AIF translocation. Clin Chim Acta. 2009 Mar;401(1-2):76-80.

[6].The selective Rho-kinase inhibitor Fasudil is protective and therapeutic in experimental autoimmune encephalomyelitis. J Neuroimmunol. 2006 Nov;180(1-2):126-34. Epub 2006 Sep 22.

Fasudil is an isoquinoline compound with a (1,4-diazacycloheptan-1-yl)sulfonyl group substituted at the 5-position. It is a Rho kinase inhibitor, and its hydrochloride hydrate is approved for the treatment of cerebral vasospasm and cerebral ischemia. Fasudil possesses a variety of pharmacological effects, including anti-aging, inhibition of EC 2.7.11.1 (a nonspecific serine/threonine protein kinase), vasodilation, nootropic effects, neuroprotection, hypotensive effects, and calcium channel blockade. It is an N-sulfonyldiazacycloheptanyl compound belonging to the isoquinoline class. It is the conjugate base of fasudil (1+). Fasudil has been investigated in the treatment of carotid artery stenosis. Introduction: Rho kinase (ROCK) plays a crucial role in the organization of the actin cytoskeleton and is involved in a variety of essential cellular functions, such as contraction and gene expression. Fasudil, a ROCK inhibitor, has been used in Japan since 1995 for the treatment of subarachnoid hemorrhage (SAH). Mounting evidence suggests that fasudil may have significant therapeutic potential for central nervous system (CNS) diseases, such as Alzheimer's disease. This article summarizes the evidence supporting the potential efficacy of fasudil in treating various CNS diseases and outlines the characteristics of its analogues. Expert opinion: Current therapies for CNS diseases only alleviate symptoms and cannot slow or halt disease progression, thus new approaches with disease-modifying effects are urgently needed. Fasudil's significant efficacy in animal models and/or clinical applications of CNS diseases makes it a promising strategy for treating human CNS diseases. Given the complex pathological mechanisms of CNS diseases, further efforts are needed to develop multifunctional fasudil derivatives or in combination with other drugs to achieve greater efficacy and minimize adverse reactions in combating CNS diseases. Blood-brain barrier (BBB) and blood-spinal cord barrier (BSCB) dysfunction are key features of multiple sclerosis (MS). We evaluated the protective effect of the selective ROCK inhibitor fasudil in a guinea pig spinal cord-induced experimental autoimmune encephalomyelitis (EAE) model. Furthermore, we investigated the effects of fasudil on the permeability of the blood-brain barrier (BBB) and blood-spinal barrier (BSCB). We found that fasudil partially alleviated experimental autoimmune encephalomyelitis (EAE)-dependent injury by reducing the permeability of the BBB and BSCB. These results provide a theoretical basis for developing selective Rho kinase inhibitors as novel therapies for multiple sclerosis (MS). https://pubmed.ncbi.nlm.nih.gov/21978848/

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Background/Objective: The Rho-ROCK signaling pathway plays a crucial role in the activation of hepatic stellate cells (HSCs). We investigated the effects of the Rho kinase (ROCK) inhibitor fasudil hydrochloride hydrate (fasudil) on HSC cell growth, collagen production, and collagenase activity. Methods: TWNT-4 cells derived from rat HSCs and human HSCs were cultured to investigate stress fiber formation and α-smooth muscle actin (α-SMA) expression. Cell proliferation was detected by BrdU incorporation assay, and apoptosis was detected by TUNEL assay. Western blotting was used to detect the phosphorylation status of MAP kinase (MAPK), extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun kinase (JNK), and p38. Enzyme-linked immunosorbent assay (ELISA) and real-time quantitative PCR were used to detect the production and gene expression of type I collagen, matrix metalloproteinase-1 (MMP-1), and tissue inhibitor of metalloproteinases-1 (TIMP-1), respectively. Collagenase activity (active MMP-1) was also detected. Results: Fasudil (100 μM) inhibited cell spreading, stress fiber formation, and α-SMA expression, accompanied by cell growth inhibition, but did not induce apoptosis. Fasudil inhibited the phosphorylation of ERK1/2, JNK, and p38. Fasudil treatment inhibited the production and transcription of collagen and TIMP, stimulated the production and transcription of MMP-1, and enhanced collagenase activity. Conclusion: These findings indicate that fasudil not only inhibits cell proliferation and collagen production but also increases collagenase activity. https://pubmed.ncbi.nlm.nih.gov/15998434/

1001206-62-7

Fasudil;103745-39-7;Fasudil dihydrochloride; 203911-27-7; 105628-07-7 (HCl); 186694-02-0 (hydrochloride hydrate)

44602745

Typically exists as solid at room temperature

2

8

2

25

513

0

MVCDPGYHRHUECK-UHFFFAOYSA-N

InChI=1S/C14H17N3O2S.CH4O3S/c18-20(19,17-9-2-6-15-8-10-17)14-4-1-3-12-11-16-7-5-13(12)14;1-5(2,3)4/h1,3-5,7,11,15H,2,6,8-10H2;1H3,(H,2,3,4)

5-(1,4-diazepan-1-ylsulfonyl)isoquinoline;methanesulfonic acid

Fasudil mesilate; fasudil mesylate; Fasudil mesilate [WHO-DD]; Fasudil (mesylate); 4S5NTB667B; UNII-4S5NTB667B; 1001206-62-7; Isoquinoline, 5-((hexahydro-1H-1,4-diazepin-1-yl)sulfonyl)-, methanesulfonate (1:1);

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Typically soluble in DMSO (e.g. 10 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)