Design of a Validated Stability Indicating HPLC Method for the analysis of Patrin-2 (Lomeguatrib).

Abstract

Lomeguatrib or Patrin-2 has been formulated to work in combination with Temozolomide. It does so by blocking the action of ATase (MGMT), which repairs specific kinds of DNA damage and is particularly responsible for resistance to anti-cancer treatments. Patrin-2 acts as a pseudosubstrate inactivator of MGMT. It thus, helps Temozolomide proceed with it’s designed action, which now cannot be stopped by neither MGMT nor MMR. The degradation behaviour of Patrin-2 was studied by subjecting it to various stress conditions. A validated stability indicating high-performance liquid chromatography method was established for the analysis of the drug in the presence of its various degradation products. An acceptable separation of the drug and its degradation products was achieved on a C18 reverse phased column (Symmetry 300) using a mobile phase that consisted of 10mM TBAA, 10mM SDS and 25mM citric acid in 35:65 acetonitrile and HPLC grade water. 285nm was determined as the wavelength max for Patrin-2, however, three wavelengths (240nm, 254nm, 285nm) were selected for the study, so as to make sure, no degradation product of Patrin-2 goes undetected. The method was validated for linearity, accuracy, precision, selectivity, specificity and robustness. It proved to be linear over the range of concentration of 1-5mg/dL (n=2) with a correlation coefficient of 0.999. The RSD was found to be as low as 0.029 and 0.233 for it’s repeatability and reproducibility. The drug was found to be unstable when exposed to 0.4M of 30%H2O2. It was highly unstable in acidic condition, undergoing complete degradation in less than an hour when subjected to 0.1M H2SO4. It underwent high degradation when subjected to photolytic stress (UV radiations) or thermal stress (incubated at 50oC and 80oC). The drug was solubilised in n-methylpyrrolidone, one of the components used to tabulate it, but the results could not be confirmed, as no degradation product could be seen, but a change in shape of the peak was observed.