Effect of a Pluronic® P123 Formulation on the Nitric Oxide-Generating Drug JS-K
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O2-(2,4-dinitrophenyl)1-[(4-ethoxycarbonyl)piperazin-1-yl]diazen-1-ium-1,2-diolate] or JS-K is a nitric oxide-producing prodrug of the arylated diazeniumdiolate class with promising anti-tumor activity. JS-K has challenging solubility and stability properties. We aimed to characterize and compare Pluronic® P123-formulated JS-K (P123/JS-K) with free JS-K.
We determined micelle size, shape, and critical micelle concentration of Pluronic® P123. Efficacy was evaluated in vitro using HL-60 and U937 cells and in vivo in a xenograft in NOD/SCID IL2Rγ null mice using HL-60 cells. We compared JS-K and P123/JS-K stability in different media. We also compared plasma protein binding of JS-K and P123/JS-K. We determined the binding and Stern Volmer constants, and thermodynamic parameters.
Spherical P123/JS-K micelles were smaller than blank P123. P123/JS-K formulation was more stable in buffered saline, whole blood, plasma and RPMI media as compared to free JS-K. P123 affected the protein binding properties of JS-K. In vitro it was as efficacious as JS-K alone when tested in HL-60 and U937 cells and in vivo greater tumor regression was observed for P123/JS-K treated NOD/SCID IL2Rγ null mice when compared to free JS-K-treated NOD/SCID IL2Rγ null mice.
Pluronic® P123 solubilizes, stabilizes and affects the protein binding characteristics of JS-K. P123/JS-K showed more in vivo anti-tumor activity than free JS-K.
KEY WORDSacute myeloid leukemia (AML) glutathione JS-K nitric oxide O2-(2,4-dinitrophenyl)1-[(4-ethoxycarbonyl)piperazin-1-yl]diazen-1-ium-1,2-diolate] Pluronic® P123
Alpha 1 acid glycoprotein
Acute myeloid leukemia
Area under the curve
Critical micelle concentration
Difference in the fluorescence in the absence and presence of JS-K or P123/JS-K at the concentration Q
Fraction of initial fluorescence accessible to quencher
Fluorescence in the absence of external quencher
High performance liquid chromatography
Human serum albumin
Association binding constant
Quenching rate constant
Stern Volmer Constant
Conc. of quencher/drug
Temperature in Kelvin
Transmission electron microscopy
Ultra performance liquid chromatography
Average lifetime of HSA molecule in absence of JS-K (quencher) or any other drug
ACKNOWLEDGMENTS AND DISCLOSURES
This work was supported by grant RO1 CA129611 from the National Cancer Institute.
Paul Shami is Scientific Founder, Chief Medical Officer and Chairman of the Board of Directors of JSK Therapeutics Inc.
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