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Alleviation of behavioral hypersensitivity in mouse models of inflammatory pain with two structurally different casein kinase 1 (CK1) inhibitors.

Kurihara T, Sakurai E, Toyomoto M, Kii I, Kawamoto D, Asada T, Tanabe T, Yoshimura M, Hagiwara M, Miyata A - Mol Pain (2014)

Bottom Line: However, the functional role of CK1 in somatosensory pain signaling has not yet been fully understood.TG003, which was originally identified as a cdc2-like kinase inhibitor, had potent inhibitory effects on CK1 isoforms in vitro and in cultured cells.Intrathecal injection of either TG003 (1-100 pmol) or IC261 (0.1-1 nmol) dose-dependently decreased mechanical allodynia and thermal hyperalgesia induced by carrageenan or CFA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pharmacology, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima City, Kagoshima 890-8544, Japan. tkmphm10@m.kufm.kagoshima-u.ac.jp.

ABSTRACT

Background: The phylogenetically highly conserved CK1 protein kinases consisting of at least seven isoforms form a distinct family within the eukaryotic protein kinases. CK1 family members play crucial roles in a wide range of signaling activities. However, the functional role of CK1 in somatosensory pain signaling has not yet been fully understood. The aim of this study was to clarify the role of CK1 in the regulation of inflammatory pain in mouse carrageenan and complete Freund's adjuvant (CFA) models.

Results: We have used two structurally different CK1 inhibitors, TG003 and IC261. TG003, which was originally identified as a cdc2-like kinase inhibitor, had potent inhibitory effects on CK1 isoforms in vitro and in cultured cells. Intrathecal injection of either TG003 (1-100 pmol) or IC261 (0.1-1 nmol) dose-dependently decreased mechanical allodynia and thermal hyperalgesia induced by carrageenan or CFA. Bath-application of either TG003 (1 μM) or IC261 (1 μM) had only marginal effects on spontaneous excitatory postsynaptic currents (sEPSCs) recorded in the substantia gelatinosa neurons of control mice. However, both compounds decreased the frequency of sEPSCs in both inflammatory pain models.

Conclusions: These results suggest that CK1 plays an important pathophysiological role in spinal inflammatory pain transmission, and that inhibition of the CK1 activity may provide a novel strategy for the treatment of inflammatory pain.

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Related in: MedlinePlus

TG003 suppresses kinase activity of CK1 family members in vitro. Recombinant CK1 family members were incubated with the substrate peptide CKtide in the presence of different concentrations of TG003 and IC261. TG003 (open circle) and IC261 (closed square) inhibited CK1 family members in a dose-dependent manner. The IC50 values of TG003 on the kinase activity of CK1α, δ, ϵ, γ1, γ2 and γ3were 0.33, 0.34, 1.4, 1.5, 0.93 and 0.88 μM, respectively. The IC50 values of IC261 were 0.19, 0.60, 0.86, > 30, > 30 and > 30 μM, respectively. Representative dose-response curves with Hill slope are shown. The results are presented as an average of duplicated experiments.
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Figure 1: TG003 suppresses kinase activity of CK1 family members in vitro. Recombinant CK1 family members were incubated with the substrate peptide CKtide in the presence of different concentrations of TG003 and IC261. TG003 (open circle) and IC261 (closed square) inhibited CK1 family members in a dose-dependent manner. The IC50 values of TG003 on the kinase activity of CK1α, δ, ϵ, γ1, γ2 and γ3were 0.33, 0.34, 1.4, 1.5, 0.93 and 0.88 μM, respectively. The IC50 values of IC261 were 0.19, 0.60, 0.86, > 30, > 30 and > 30 μM, respectively. Representative dose-response curves with Hill slope are shown. The results are presented as an average of duplicated experiments.

Mentions: We first verified the inhibitory effect of TG003 on the enzymatic activity of CK 1 family members and compared it with that of IC261. Recombinant CK1α, δ, ϵ, γ1, γ2 or γ3 were incubated with the substrate peptide CKtide in the presence of different concentrations of TG003 or IC261, respectively. Both small molecules inhibited CK1 family members in a dose-dependent manner (Figure 1). Inhibition of CK1α, δ and ϵ by TG003 were equivalent to that by IC261. On the other hand, the inhibitory activities of TG003 on the kinase activity of CK1γ1, γ2 and γ3 were 20-fold or more higher than those of IC261 (Figure 1). These results indicate that TG003 and IC261 are able to suppress the kinase activity of broad CK1 family members equally.


Alleviation of behavioral hypersensitivity in mouse models of inflammatory pain with two structurally different casein kinase 1 (CK1) inhibitors.

Kurihara T, Sakurai E, Toyomoto M, Kii I, Kawamoto D, Asada T, Tanabe T, Yoshimura M, Hagiwara M, Miyata A - Mol Pain (2014)

TG003 suppresses kinase activity of CK1 family members in vitro. Recombinant CK1 family members were incubated with the substrate peptide CKtide in the presence of different concentrations of TG003 and IC261. TG003 (open circle) and IC261 (closed square) inhibited CK1 family members in a dose-dependent manner. The IC50 values of TG003 on the kinase activity of CK1α, δ, ϵ, γ1, γ2 and γ3were 0.33, 0.34, 1.4, 1.5, 0.93 and 0.88 μM, respectively. The IC50 values of IC261 were 0.19, 0.60, 0.86, > 30, > 30 and > 30 μM, respectively. Representative dose-response curves with Hill slope are shown. The results are presented as an average of duplicated experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4008364&req=5

Figure 1: TG003 suppresses kinase activity of CK1 family members in vitro. Recombinant CK1 family members were incubated with the substrate peptide CKtide in the presence of different concentrations of TG003 and IC261. TG003 (open circle) and IC261 (closed square) inhibited CK1 family members in a dose-dependent manner. The IC50 values of TG003 on the kinase activity of CK1α, δ, ϵ, γ1, γ2 and γ3were 0.33, 0.34, 1.4, 1.5, 0.93 and 0.88 μM, respectively. The IC50 values of IC261 were 0.19, 0.60, 0.86, > 30, > 30 and > 30 μM, respectively. Representative dose-response curves with Hill slope are shown. The results are presented as an average of duplicated experiments.
Mentions: We first verified the inhibitory effect of TG003 on the enzymatic activity of CK 1 family members and compared it with that of IC261. Recombinant CK1α, δ, ϵ, γ1, γ2 or γ3 were incubated with the substrate peptide CKtide in the presence of different concentrations of TG003 or IC261, respectively. Both small molecules inhibited CK1 family members in a dose-dependent manner (Figure 1). Inhibition of CK1α, δ and ϵ by TG003 were equivalent to that by IC261. On the other hand, the inhibitory activities of TG003 on the kinase activity of CK1γ1, γ2 and γ3 were 20-fold or more higher than those of IC261 (Figure 1). These results indicate that TG003 and IC261 are able to suppress the kinase activity of broad CK1 family members equally.

Bottom Line: However, the functional role of CK1 in somatosensory pain signaling has not yet been fully understood.TG003, which was originally identified as a cdc2-like kinase inhibitor, had potent inhibitory effects on CK1 isoforms in vitro and in cultured cells.Intrathecal injection of either TG003 (1-100 pmol) or IC261 (0.1-1 nmol) dose-dependently decreased mechanical allodynia and thermal hyperalgesia induced by carrageenan or CFA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pharmacology, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima City, Kagoshima 890-8544, Japan. tkmphm10@m.kufm.kagoshima-u.ac.jp.

ABSTRACT

Background: The phylogenetically highly conserved CK1 protein kinases consisting of at least seven isoforms form a distinct family within the eukaryotic protein kinases. CK1 family members play crucial roles in a wide range of signaling activities. However, the functional role of CK1 in somatosensory pain signaling has not yet been fully understood. The aim of this study was to clarify the role of CK1 in the regulation of inflammatory pain in mouse carrageenan and complete Freund's adjuvant (CFA) models.

Results: We have used two structurally different CK1 inhibitors, TG003 and IC261. TG003, which was originally identified as a cdc2-like kinase inhibitor, had potent inhibitory effects on CK1 isoforms in vitro and in cultured cells. Intrathecal injection of either TG003 (1-100 pmol) or IC261 (0.1-1 nmol) dose-dependently decreased mechanical allodynia and thermal hyperalgesia induced by carrageenan or CFA. Bath-application of either TG003 (1 μM) or IC261 (1 μM) had only marginal effects on spontaneous excitatory postsynaptic currents (sEPSCs) recorded in the substantia gelatinosa neurons of control mice. However, both compounds decreased the frequency of sEPSCs in both inflammatory pain models.

Conclusions: These results suggest that CK1 plays an important pathophysiological role in spinal inflammatory pain transmission, and that inhibition of the CK1 activity may provide a novel strategy for the treatment of inflammatory pain.

Show MeSH
Related in: MedlinePlus