EHT 1864 2HCl (EHT-1864; EHT1864), the dihydrochloride salt of EHT-1864, is a potent and novel Rac family GTPase inhibitor with important biological activity. It inhibits Rac1, Rac1b, Rac2 and Rac3 with Kd values of 40 nM, 50 nM, 60 nM and 250 nM, respectively. EHT 1864 attenuates glucose-stimulated insulin secretion in pancreatic β-cells. EHT 1864 specifically inhibits glucose-induced Rac1 activation and membrane association and associated downstream signaling events culminating in inhibition of GSIS.
理化性质和储存条件
| Molecular Weight (MW) | 581.47 |
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| Formula | C25H29Cl2F3N2O4S |
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| CAS No. | 754240-09-0 |
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| Storage | -20℃ for 3 years in powder form |
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| -80℃ for 2 years in solvent |
| Solubility (In vitro) | DMSO: 100 mg/mL (172.0 mM) |
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| Water: 100 mg/mL (172.0 mM) |
| Ethanol: <1 mg/mL |
| SMILES | O=C1C=C(CN2CCOCC2)OC=C1OCCCCCSC3=CC=NC4=CC(C(F)(F)F)=CC=C34 |
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| Synonyms | EHT-1864; EHT1864; EHT 1864 HCl |
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实验参考方法
| In Vitro | In vitro activity: EHT 1864 selectively inhibits Rac-induced lamellipodia formation, and specifically reverses cell transformation induced by constitutively activated mutants of Rac1 and Tiam1. EHT 1864 strongly impaires oncogenic Ras-induced cell proliferation in NIH 3T3 cells stably expressing H-Ras(61L) protein. EHT 1864 also reduces both extracellular and intracellular levels of Aβ peptides by inhibiting the γ-secretase-dependent cleavage of APP. In cultured hippocampal pyramidal neurons, EHT 1864, via inhibition of Rac1, rescues the phenotype induced by Rich2 knock-down.
Kinase Assay: For inhibitor:GTPase binding analyses, aliquots of small GTPase solution (containing 1 μM inhibitor) are titrated into a solution of 1 μM inhibitor in the cuvette. Changes in fluorescence anisotropy are monitored at λex = 360 nm, λem = 440 nm, 30 s after each addition. All data analysis and curve fitting were performed using Microsoft Excel and QuantumSoft's ProFit for Mac OS X.
Cell Assay: NIH 3T3 cells stably expressing oncogenic Ras are plated in 96-well plates. The cells are cultured for up to 4 days in complete growth medium, either alone, or supplemented with 5 μM EHT 1864. Cell growth is then assessed using the conversion of MTT to a formazan product. Briefly, the MTT reagent (from a 5 mg/ml solution diluted in PBS) is added to the wells at a final concentration of 0.5 mg/ml, and the cells are further incubated for 4 h at 37°C. The medium is then removed, and the reaction is terminated by adding 100 μl/well Me2SO. The absorbance is read at 570 nm using a microplate reader. |
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| In Vivo | EHT 1864 (40 mg/kg i.p.) significantly reduces Abeta 40 and Abeta 42 levels in guinea pig brains. |
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| Animal model | Male Hartley albino guinea pigs |
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| Formulation & Dosage | Dissolved in saline; 40 mg/kg daily; i.p. injection |
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| References | J Biol Chem. 2007 Dec 7;282(49):35666-78; J Biol Chem. 2005 Nov 11;280(45):37516-25. |
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