Experimental and Computational Evaluation of Chloranilic Acid as an Universal Chromogenic Reagent for the Development of a Novel 96-Microwell Spectrophotometric Assay for Tyrosine Kinase Inhibitors

Abstract

The tyrosine kinase inhibitors (TKIs) are chemotherapeutic drugs used for the targeted therapy of various types of cancer. This work discusses the experimental and computational evaluation of chloranilic acid (CLA) as a universal chromogenic reagent for developing a novel 96-microwell spectrophotometric assay (MW-SPA) for TKIs. The reaction resulted in an instantaneous formation of intensely purple colored products with TKIs. Spectrophotometric results confirmed that the reactions proceeded via the formation of charge-transfer complexes (CTCs). The physical parameters were determined for the CTCs of all TKIs. Computational calculations and molecular modelling for the CTCs were conducted, and the site(s) of interaction on each TKI molecule were determined. Under the optimized conditions, Beer’s law correlating the absorbances of the CTCs with the concentrations of TKIs were obeyed in the range of 10–500 µg/well with good correlation coefficients (0.9993–0.9998). The proposed MW-SPA fully validated and successfully applied for the determination of all TKIs in their bulk forms and pharmaceutical formulations (tablets). The proposed MW-SPA is the first assay that can analyze all the TKIs on a single assay system without modifications in the detection wavelength. The advantages of the proposed MW-SPA are simple, economic and, more importantly, have high throughput.

Synthesis, spectroscopic and computational characterization of charge transfer complex of remdesivir with chloranilic acid: Application to development of novel 96-microwell spectrophotometric assay

“…As a result, these assays are costly, and they also expose the analyst to the harmful impacts of organic solvents [ 53 , 54 ]. Darwish et al [21] , [22] , [23] , [24] , [25] , [26] , [27] , [28] , [29] , [30] previously developed a number of successful microwell-based photometric methods to quantify the active drug contents in pharmaceutical dosage forms using an absorbance plate reader. These methods have a high throughput and use small amounts of organic solvents.…”

“…CTC creation is also crucial in a multitude of fields, including the production of optical and electrically conductive materials [10] , [11] , [12] , [13] , microemulsions and surface chemistry [14] , photocatalysis [15] , dendrimers [16] , solar energy storage [17] , and organic semiconductors [18] . Additionally, the generation of CTC between drugs and specific σ- and π-acceptors has been effectively employed as the basis for designing high-accuracy assays for both qualitative and quantitative detection of pharmaceuticals in bulk and/or pharmaceutical dose forms [19] , [20] , [21] , [22] , [23] , [24] , [25] , [26] , [27] , [28] , [29] , [30] .…”

“…We've been evaluating the development and applications of CTC in a variety of pharmaceuticals with diverse acceptors over the past few years. Antibiotics, kinase inhibitors, antihypertensives and antihyperlipidemics were among the pharmaceuticals used [19] , [20] , [21] , [22] , [23] , [24] , [25] , [26] , [27] , [28] , [29] , [30] . Our research focused on understanding the chemistry of these pharmaceuticals' CT reactions and their electron donating manners toward a variety of electron acceptors.…”

Pretreatment and analysis techniques development of TKIs in biological samples for pharmacokinetic studies and therapeutic drug monitoring

“…Conventional spectrophotometric methods, such as UV spectrophotometry based on conjugated structure and fluorescence spectrophotometry based on fluorescence properties [ 136 , 137 ], are simple and sensitive techniques for the detection of TKIs. Assays based on ion-pair associates [ 138 ] and 96-microwell-based spectrophotometric assay [ 139 , 140 ] have been reported for the convenient and accurate determination of TKIs in pharmaceutical formulations. However, traditional spectrophotometric methods may face challenges in distinguishing certain TKI structures (i.e., vandetanib, dasatinib, and sorafenib) due to their similar UV-visible characteristics [ 141 ].…”

Development of Green and High Throughput Microplate Reader-Assisted Universal Microwell Spectrophotometric Assay for Direct Determination of Tyrosine Kinase Inhibitors in Their Pharmaceutical Formulations Irrespective the Diversity of Their Chemical Structures

“…For these reasons, the development of a universal spectrophotometric assay for quantitation of all TKIs irrespective of their different chemical structure, and overcoming these drawbacks, would be very valuable. In a previous study, Darwish et al [ 51 ] developed a universal spectrophotometric assay for TKIs; however, the assay is still ever suffering from the use of health/environment hazardous derivatizing reagents.…”