Drug development and bioanalytical method validation for a novel anticancer molecule, 4‐(dimethylamino)‐2‐(p‐tolylamino) thiazole‐5‐carbonitrile

Abstract

Hit, Lead & Candidate Discovery The thiazole ring system represents a significant building block that exists in many biologically active natural products and clinically successful anticancer drugs. Modifications of the thiazole core have been a proven and highly effective method in improving anticancer potency. We designed a novel thiazole‐based molecule, 4‐(dimethylamino)‐2‐(p‐tolylamino) thiazole‐5‐carbonitrile, which showed potent in vitro anticancer effect against targeted Bcl‐2 Jurkat cell‐line quantified using 3‐(4, 5‐dimethythiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide (MTT) assay. Physicochemical parameters (pKa, LogP, LogD) of the molecule have also been evaluated as a part of the drug development process. Moreover, a rapid Reverse phase‐high performance liquid chromatography (RP‐HPLC) bioanalytical method has been developed to quantitate the molecule in human plasma. The method has been validated following the United States Food and Drug Administration bioanalytical method validation guideline. The stability studies showed no significant instability of the analyte in respective stability conditions (6 hr autosampler, 8 hr bench top, 30 days long‐term, and 3 freeze–thaw cycles). The molecule was found to be stable for 3 hr in human plasma. The molecule was shown to have high plasma protein binding affinity. It showed favorable pKa (11), Log P (3.01), Log D (2.96), and plasma protein binding (90%) values toward its further exploitation as a lead anticancer candidate molecule. The developed bioanalytical method can be used for quantifying the molecule in different pharmacokinetic, toxicokinetic, or other clinical trial samples involving human plasma during development process or in routine bioavailability and bioequivalence study after its regulatory approval.

Impact of Disease States on the Oral Pharmacokinetics of EIDD-1931 (an Active Form of Molnupiravir) in Rats for Implication in the Dose Adjustment

“…The chemical stability of EIDD-1931 was estimated in simulated gastric fluid (SGF) using omeprazole as the standard (Tables S2 and S5). We used earlier reported in-house protocols to assess metabolic stability, , ionization constant, , plasma protein binding, and chemical stability. , The detailed methodologies are described in the Supporting Information.…”

Development of a Simple, Precise, and Validated HPLC Method for the Anticancer Drug, Regorafenib: Application to Pharmacokinetics in Rats and Stability Study

“…For the second storage condition, three cycles of repeated freeze and thaw were carried out before the assessment of the samples. Each cycle consists of 2–3 h thawing at room temperature and freezing for 24 h 13 . The third storage condition involved keeping frozen samples (−20 °C) at autosampler for 48 h before the HPLC evaluation 14 …”

Paradigm Shift in the Arena of Sample Preparation and Bioanalytical Approaches Involving Liquid Chromatography Mass Spectroscopic Technique

“…66 Protein precipitation has been extensively used to extract the analyte from biological matrices over the years in pharmacokinetic studies. 67,68 Quantification of bioactive flavonoid, alpinetin in rat plasma using UHPLC-MS/ MS has been carried out by Ye et al Protein precipitation together with dilution approach using acetonitrile and water (50:50, v/v) was used to eliminate the matrix components. 69 Mohammed et al published a fast and sensitive validated bioanalytical method for quantification of Linezolid using a simple protein extraction procedure with methanol.…”

“…These techniques are reported to be used for sample extraction in pharmacokinetics studies, biomarker discovery, and pharmacodynamic studies. 68,87,91 Capka and Carter reported the development and validation of an LC-MS/MS method for determining salmeterol in human plasma using mixed mode anion exchange SPE for sample clean up combined with the column switching approach to reduce matrix interference. Salmeterol is a basic drug having pKa about 9.3.…”