Cell lines

Human hepatic stellate cell line LX-2

Preparation method

Soluble in DMSO >30mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

5, 10, 25, 50μmol/L for 24h; 10 μmol/L for 4, 8, 16, 24, 48h

Applications

KN-93 decreased the proliferation of human hepatic stellate cells in a dose dependent manner after 24 h treatment. Incubation of 10 μmol/L KN-93 induced the cell growth reduction in a time-dependent manner. KN-93 reduced the expression of p53 and p21.

Animal models

adult Sprague Dawley female rats (model of PD was induced by 6-hydroxydopamine (OHDA))

Dosage form

1μg, 2μg, or 5μg KN-93, intrastriatal injections

Application

The data indicated that intrastriatal injections of KN-93 were beneficial in reducing the expression of LID (levodopa-induced dyskinesia) by lowering the expression of pGluR1S845(Phosphorylated GluR1(Glutamate receptor 1) at Serine-845) via suppressing the activation of CaMKII (calcium -dependent protein kinase II) in PD rats. Decreased expression of pGluR1S845 further reduced the expression of Gad1(glutamate decarboxylase 1) and nr4a1(nuclear receptor subfamily 4, group A, member 1) in PD rats.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

KN-93 phosphate is the water soluble form of KN-93. It is a potent, selective and cell permeable inhibitor of CaM kinase II (IC50= 0.37 μm, Ki=370 nm) [1]
CaMK II (calcium/calmodulin-dependent protein kinase II) is a multifunctional serine/threonine kinase with various roles in different physiological systems.
Since CaMK II can enhance cell proliferation, the CaMK II inhibitor KN-93 blocked the cell proliferation by regulating the expression of p53 and p21 in human hepatic stellate cells. [2] KN-93 inhibited the autophosphorylation of both α and β subunits of CaMK II. Moreover, KN-93 reduced dopamine level via modulating the TH reaction rate to reduce the Ca2+-mediated phosphorylation levels of the TH molecule in PC12h cells.[1] As CaMK II is necessary for cell cycle progression through G1, KN-93 also induced G1 cell cycle arrest and apoptosis in NIH 3T3 cells.[3]
In MRL/lpr Foxp3-GFP mice, KN-93 promoted the generation of Foxp3 regulatory T cells through the inhibition of CaMK IV in the spleen, peripheral lymph nodes and peripheral blood with decreased skin and kidney damage. [4] In a rat model of Parkinson's disease, intrastriatal injection of KN-93 relieved the levodopa-induced dyskinesia by suppressing the CaMK II activation. [5]
References:
[1] Sumi M, Kiuchi K, Ishikawa T, Ishii A, Hagiwara M, Nagatsu T, Hidaka H. The newly synthesized selective Ca2+/calmodulin dependent protein kinase II inhibitor KN-93 reduces dopamine contents in PC12h cells. Biochem Biophys Res Commun. 1991 Dec 31;181(3):968-75.
[2] An P, Zhu JY, Yang Y, Lv P, Tian YH, Chen MK, Luo HS. KN-93, a specific inhibitor of CaMKII inhibits human hepatic stellate cell proliferation in vitro. World J Gastroenterol. 2007 Mar 7;13(9):1445-8.
[3] Tombes RM, Grant S, Westin EH, Krystal G. G1 cell cycle arrest and apoptosis are induced in NIH 3T3 cells by KN-93, an inhibitor of CaMK-II (the multifunctional Ca2+/CaM kinase). Cell Growth Differ. 1995 Sep;6(9):1063-70.
[4] Koga T, Mizui M, Yoshida N, Otomo K, Lieberman LA, Crispín JC, Tsokos GC.KN-93, an inhibitor of calcium/calmodulin-dependent protein kinase IV, promotes generation and function of Foxp3+ regulatory T cells in MRL/lpr mice. Autoimmunity. 2014 Nov;47(7):445-50.
[5] Yang X, Wu N, Song L, Liu Z. Intrastriatal injections of KN-93 ameliorates levodopa-induced dyskinesia in a rat model of Parkinson's disease. Neuropsychiatr Dis Treat. 2013;9:1213-20. doi: 10.2147/NDT.S45422.