Intranasal delivery of FSD‐C10, a novel Rho kinase inhibitor, exhibits therapeutic potential in experimental autoimmune encephalomyelitis

Abstract

SummaryViewing multiple sclerosis (MS) as both neuroinflammation and neurodegeneration has major implications for therapy, with neuroprotection and neurorepair needed in addition to controlling neuroinflammation in the central nervous system (CNS). While Fasudil, an inhibitor of Rho kinase (ROCK), is known to suppress experimental autoimmune encephalomyelitis (EAE), an animal model of MS, it relies on multiple, short-term injections, with a narrow safety window. In this study, we explored the therapeutic effect of a novel ROCK inhibitor FSD-C10, a Fasudil derivative, on EAE. An important advantage of this derivative is that it can be used via non-injection routes; intranasal delivery is the preferred route because of its efficient CNS delivery and the much lower dose compared with oral delivery. Our results showed that intranasal delivery of FSD-C10 effectively ameliorated the clinical severity of EAE and CNS inflammatory infiltration and promoted neuroprotection. FSD-C10 effectively induced CNS production of the immunoregulatory cytokine interleukin-10 and boosted expression of nerve growth factor and brain-derived neurotrophic factor proteins, while inhibiting activation of p-nuclear factor-jB/p65 on astrocytes and production of multiple pro-inflammatory cytokines. In addition, FSD-C10 treatment effectively induced CD4 + CD25 + , CD4 + FOXP3 + regulatory T cells. Together, our results demonstrate that intranasal delivery of the novel ROCK inhibitor FSD-C10 has therapeutic potential in EAE, through mechanisms that possibly involve both inhibiting CNS inflammation and promoting neuroprotection.

FSD-C10, a Fasudil derivative, promotes neuroregeneration through indirect and direct mechanisms

“…Similar to our previous study19, nasal administration of FSD-C10 effectively suppressed clinical severity of EAE, with reduced CNS inflammation and demyelination (Figure S1). Extensive CD4 + T cells and CD68 + macrophages were found in brains from untreated EAE mice whereas the frequency of these cells were significantly reduced in mice treated with nasal FSD-C10 (Figure S2).…”

“…Inhibition of Rho associated kinase (ROCK) has been shown to promote OPC differentiation and myelin formation29303132. We have recently found that nasal administration of FSD-C10, a novel and efficient ROCK inhibitor, suppressed the activity and expression of ROCK, alleviated disease severity and CNS inflammation of EAE mice, and preserved myelin sheath1933. Our data showed a higher proliferation rate of OPCs and mature oligodendrocytes in the CNS of FSD-C10-treated mice, which confirms the protective effect of FSD-C10 on CNS myelination in vivo .…”

“…EAE were induced as described previously19. Mice were subcutaneously immunized with 300 μg mouse myelin oligodendrocyte glycoprotein 35–55 peptide (MOG 35–55 , MEVGWYRSPFSRVVHLYRNGK) in Freund’s complete adjuvant (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 3 mg/ml of Tuberculosis H37Ra (BD Difco, NJ, USA).…”

“…In this context, our previous study showed that nasal administration of FSD-C10, a derivative of Fasudil with less toxic effect, effectively suppressed the clinical severity of experimental autoimmune encephalomyelitis (EAE), an animal model of MS. This effect was associated with a upregulated Tregs19. Still, whether FSD-C10 presents a neuroregenerative and neuroprotective effect has yet to be elucidated.…”

The role of growth factors as a therapeutic approach to demyelinating disease

“…Successful delivery of BDNF, CNTF (Alcala-Barraza et al 2010, Vaka et al 2012), IGF-I (Thorne et al 2004), and FGF-2 (Feng et al 2012) is reported in rodents. Intranasal administration of proteins is also used in humans (Reger et al 2006) and in EAE models of MS (Li et al 2014). …”