Equilibria, kinetics and mechanism for the degradation of the cytotoxic compound L-N^G-nitroarginine

Abstract

L-N^G-nitroarginine (LNNA), an analog of L-arginine, is a competitive inhibitor of nitric oxide synthase which causes the selective reduction of blood flow to tumor cells. Despite the potential of LNNA to function as an adjuvant in cancer therapies, its poor solubility and stability have hindered the development of an injectable formulation of LNNA that is suitable for human administration. This work, for the first time, details a systematic study on the determination of equilibrium K_a constants and the rate law of LNNA degradation. The four K_a values of LNNA were determined to be 1.03, 1.10 × 10⁻², 2.51 × 10⁻¹⁰, and 1.33 × 10⁻¹³ M. From the kinetic and equilibrium studies, we have shown that the deprotonated form of LNNA is the main form of LNNA that undergoes degradation in aqueous media at room temperature. The rate law of LNNA degradation was found to be first order with respect to OH⁻ concentration and first order with respect to LNNA⁻ concentration. The rate constant at 25 °C and 1 atm was determined to be 0.04453 M⁻¹min⁻¹. A base catalyzed mechanism of LNNA degradation was proposed based on the kinetic study. The mechanism was found to be very useful in explaining the discrepancies and changes of the rate law at different pH values. It is thus recommended that LNNA should be formulated as a concentrated solution in acidic conditions for maximum chemical stability during storage and be diluted with a basic solution to near physiological pH just before administration.

Keywords:

• L-N^G-nitroarginine
• solubility enhancement
• equilibrium constants
• degradation kinetics
• rate law

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

The authors would like to acknowledge the financial support from GEA-NUS PPRL fund (N-148–000-008–001) and grant NMRC/CIRG/1339/2012.