Identification and characterization of the rat in-vivo and in-vitro metabolites of tazemetostat using LC-QTOF-MS.

In drug discovery, metabolite profiling unveils biotransformation pathways and potential toxicant formation, guiding selection of candidates with optimal pharmacokinetics and safety profiles. Tazemetostat (TAZ) is employed in treating locally advanced or metastatic epithelioid sarcoma. Identification of drug metabolites are of significant importance in improving safety, efficacy and reduced toxicity of drugs.

Induction effect of antiretroviral bictegravir on the expression of Abcb1, Abcg2 and Abcc1 genes associated with P-gp, BCRP and MRP1 transporters present in rat peripheral blood mononuclear cells.

Background: Antiretrovirals have the potential to cause drug interactions leading to inefficacy or toxicity via induction of efflux transporters through nuclear receptors, altering drug concentrations at their target sites.

Research Design And Methods: This study used molecular dynamic simulations and qRT-PCR to investigate bictegravir's interactions with nuclear receptors PXR and CAR, and its effects on efflux transporters (P-gp, BCRP, MRP1) in rat PBMCs. PBMC/plasma drug concentrations were measured using LC-MS/MS to assess the functional impact of transporter expression.

Development of an effective cleaning technique and ancillary analytical method for estimation of residues of selected kinase inhibitors from stainless steel and glass surfaces by swab sampling.

Importance of cleaning validation in the pharmaceutical industry cannot be overstated. It is essential for preventing cross-contamination, ensuring product quality & safety, and upholding regulatory standards. The present study involved development of an effective cleaning method for five selected kinase inhibitors binimetinib (BMT), selumetinib (SMT), brigatinib (BGT), capmatinib (CPT), and baricitinib (BRT).

Ultra-high-performance liquid chromatography-quadrupole time of flight tandem mass spectrometry based in vitro metabolite profiling of DK-GV-04P, a novel anticancer molecule under drug discovery.

DK-GV-04P, chemically identified as 3-cinnamyl-2-(4-methoxyphenyl) quinazolin-4(3H)-one, is an investigational molecule synthesized at the Chemical Biology Laboratory of the National Institute of Pharmaceutical Education and Research-Ahmedabad. The compound has shown potential anticancer activity against squamous CAL27 cell lines. Metabolite identification and characterization are critical in drug discovery, providing key insights into a compound's pharmacokinetics, pharmacodynamics safety, and metabolic fate.

Comprehensive characterization and preclinical assessment of an imidazopyridine-based anticancer lead molecule.

Imidazopyridine scaffold holds significant pharmacological importance in the treatment of cancer. An in-house synthesized imidazopyridine-based molecule was found to have promising anticancer activity against breast cancer, lung cancer, and colon cancer. The molecule is an inhibitor of pyruvate kinase M2, the enzyme that elevates tumor growth, metastasis and chemoresistance by directly controlling tumor cell metabolism.

Delineation of prototypical degradation mechanism, characterization of unknown degradation impurities by liquid chromatography-quadrupole-time-of-flight-tandem mass spectrometry and stability-indicating analytical method of selumetinib.

Selumetinib (SELU) was recently approved by the US Food and Drug Administration (US FDA) in 2020. However, the degradation impurities of SELU have not been characterized or identified to date. The mechanism for impurity formation and the degradation behavior have not been previously studied.

Differential role of potential stressors, underlying degradation mechanism, characterization of degradants using LC-MS/MS, and establishment of a stability-indicating analytical method for duvelisib.

Duvelisib (DUV) was first approved globally in 2018. An extensive literature search revealed that the differential role of a potential degradation medium in altering the shelf-life of DUV due to its exposure during storage has not been identified till date. Moreover, its degradation impurities and degradation mechanism are not known.

Potential degradation products of abemaciclib: Identification and structural characterization employing LC-Q/TOF-MS and NMR including mechanistic explanation.

Degradation products are the potential drug impurities that can be generated during transport and storage of pharmaceuticals. Before this study, degradation chemistry and potential degradation products of abemaciclib (ABM) were unknown. Moreover, no stability-indicating analytical method was available that can be used to analyse ABM in presence of its degradation products.

Maresin-1 prevents blood-spinal cord barrier disruption associated with TRPV4 elevation in the experimental model of spinal cord injury.

Recently, we reported the TRPV4 ion channel activation and its association with secondary damage after spinal cord injury (SCI). TRPV4 activation is linked with blood-spinal cord barrier (BSCB) disruption, endothelial damage, and inflammation after SCI. Specialized pro-resolving mediators (SPM) are endogenous lipid mediators released for inflammation resolution.

Progressive tools and critical strategies for development of best fit PBPK model aiming better in vitro-in vivo correlation.

Nowadays, conducting discriminative dissolution experiments employing physiologically based pharmacokinetic modeling (PBPK) or physiologically based biopharmaceutical modeling (PBBM) is gaining significant importance in quantitatively predicting oral absorption of drugs. Mechanistic understanding of each process involved in drug absorption and its impact on the performance greatly facilitates designing a formulation with high confidence. Unfortunately, the biggest challenge scientists are facing in current days is the lack of standardized protocol for integrating dissolution experiment data during PBPK modeling.

Assessment of metabolic stability and pharmacokinetics by LC-MS/MS and establishment of the safe dose of IMID-2, a novel anticancer molecule under drug discovery.

Pyruvate kinase (PK) M2 activators ramp up glycolysis in cancer cells, leading to a reversal of the Warburg effect in cancer cells. A promising PKM2 activator molecule, IMID-2, developed by the National Institute of Pharmaceutical Education and Research-Ahmedabad showed promising anticancer activity against MCF-7 and COLO-205 cell lines, which represent breast and colon cancer. Its physicochemical properties, like solubility, ionization constant, partition coefficient and distribution constant, have already been established.

LC-QQQ-MS based intracellular quantification of bictegravir in peripheral blood mononuclear cells and plasma.

Most antiretrovirals (ARVs) have intracellular therapeutic target sites and therefore, their plasma concentration may be misleading when relating to their efficacy or toxicity. A bioanalytical method for quantification of the ARV drug bictegravir (BTG) in its target site peripheral blood mononuclear cells (PBMCs) is not available till date. This is the first time to establish a sufficiently sensitive mass spectrometry-based bioanalytical method to quantify BTG in both rat PBMCs and plasma.

ProteoExcel: Development of a simple excel-based tool for surrogate peptide selection in mass spectrometry based targeted proteomics.

Selection of surrogate peptides plays a major role to achieve reproducible and accurate quantification of desired proteins in targeted proteomics. Currently, available peptide selection tools suffer from the limitation of not covering entire proteins including all the species and inflexibility in applying the exclusion criteria. Here, we have developed an excel-based ProteoExcel tool which can automatically select the most appropriate surrogate peptides with high flexibility in terms of addition, deletion or changing the exclusion criteria.

Mechanism of capmatinib degradation in stress conditions including degradation product characterization using ultra-high-performance liquid chromatography-quadrupole-time of flight mass spectrometry and stability-indicating analytical method development.

Rationale: Capmatinib (CMT) has been recently approved for the treatment of non-small cell lung cancer by the United States Food and Drug Administration (USFDA). Till date, the degradation mechanism of CMT in different stress conditions is not known. Moreover, degradation products (DPs) of the drug are yet to be identified.

Bioanalysis by LC-MS/MS and preclinical pharmacokinetic interaction study of ribociclib and oleanolic acid.

Ribociclib (RIBO), approved in 2017 for HR-positive and HER-2-negative metastatic breast cancer treatment is reported to have the potential to induce hepatobiliary toxicity in patients. Oleanolic acid (OLA) has hepatoprotective potential that can be beneficial if coadministered with RIBO. The primary scope of this study was to develop quantitative bioanalytical methods for RIBO and OLA.

LC/Q-TOF MS and LC/QQQ MS based bioanalysis of a new ferrocene derivative as a potential anticancer lead with promising drug-like characteristics.

Pyrazolopyrimidine ring present in various approved drugs is reported to target the tyrosine kinase receptor. A new pyrazolopyrimidine ferrocene derivative, which targets tumor pyruvate kinase M2 showed an impressive antiproliferative profile against human oral squamous cell carcinoma cell line CAL27 assessed using Alamar blue assay. In line with the lead optimization process, the molecule was studied for physicochemical properties where a bioanalytical method has been developed in plasma on liquid chromatography-mass spectrometry and validated following the USFDA bioanalytical method validation guideline.

High resolution mass spectrometry-driven metabolite profiling of baricitinib to report its unknown metabolites and step-by-step reaction mechanism of metabolism.

Rationale: Metabolite profiling is an integral part of the drug development process for selecting candidates with high therapeutic efficacy and low risk. Baricitinib (BARI) was approved in 2018 by the US Food and Drug Administration to treat rheumatoid arthritis. According to the available literature, no systematic study has been reported on the metabolite profiling of BARI.

Surrogate peptide selection and internal standardization for accurate quantification of endogenous proteins.

Relative quantification techniques have dominated the field of proteomics. However, biomarker discovery, mathematical model development and studies on transporter-mediated drug disposition still need absolute quantification of proteins. The quality of data of trace-level protein quantification is solely dependent on the specific selection of surrogate peptides.

Degradation kinetics and characterization of major degradants of binimetinib employing liquid chromatography-high resolution mass spectrometry.

Binimetinib (BMT) has recently been approved by the USFDA for the treatment of melanomas. An extensive literature search revealed that degradation kinetics of BMT is not reported in any scientific report. Till date, no stability indicating analytical method (SIAM) is available for quantification of BMT in presence of its impurities.

Implication of metabolomics and transporter modulation based strategies to minimize multidrug resistance and enhance site-specific bioavailability: a needful consideration toward modern anticancer drug discovery.

Induction of drug-metabolizing enzymes and efflux transporters (DMET) through activation of pregnane x receptor (PXR) is the primary factor involved in almost all bioavailability and drug resistance-related problems of anticancer drugs. PXR is a transcriptional regulator of many metabolizing enzymes and efflux transporters proteins like p-glycoprotein (p-gp), multidrug resistant protein 1 and 2 (MRP 1 and 2), and breast cancer resistant protein (BCRP), etc. Several anticancer drugs are potent activators of PXR receptors and can modulate the gene expression of DMET proteins.

Advancement in Analytical Strategies for Quantification of Biomarkers with a Special Emphasis on Surrogate Approaches.

Accurate quantification of biomarkers has always been a challenge for many bioanalytical scientists due to their endogenous nature and low concentration in biological matrices. Different analytical approaches have been developed for quantifying biomarkers including enzyme-linked immunosorbent assay, immunohistochemistry, western blotting, and chromatographic techniques assisted with mass spectrometry. Liquid chromatography-tandem mass spectrometry-based quantification of biomarkers has gained more attention over other traditional techniques due to its higher sensitivity and selectivity.

UHPLC-Q-TOF-MS/MS-based metabolite profiling of duvelisib and establishment of its metabolism mechanisms.

Duvelisib is a dual inhibitor of phosphoinositide 3 kinase that received global approval by the US Food and Drug Administration in 2018 to treat follicular lymphoma after at least two prior systemic therapies. An extensive literature search revealed that, to date, metabolites of duvelisib have not been characterized and information on them is not available in any of the literature. Moreover, the metabolism pathway is yet to be established.

LC-MS/MS bioanalytical method for quantification of binimetinib and venetoclax, and their pharmacokinetic interaction.

Because of several prospective benefits, binimetinib (BMT)-venetoclax (VTC) combination can be a better therapeutic strategy to treat cancer. An LC-MS/MS method for simultaneous quantification of BMT and VTC in rat plasma has been developed and validated. Specificity, accuracy, precision and stability results met the acceptance criteria for validation.

Innovation in Strategies for Sensitivity Improvement of Chromatography and Mass Spectrometry Based Analytical Techniques.

Chromatography and mass spectrometry based techniques are the most commonly employed procedures to quantitate the analytes in pharmaceutical research. However, sensitivity of analytical methods significantly varies due to the difference in physicochemical characteristics of analytes. Sensitivity of methods greatly affects the quality of analytical results.

Advancements in practical and scientific bioanalytical approaches to metabolism studies in drug development.

Advancement in metabolism profiling approaches and bioanalytical techniques has been revolutionized over the last two decades. Different and approaches along with advanced bioanalytical techniques are enabling the accurate qualitative and quantitative analysis of metabolites. This review summarizes various modern and approaches for executing metabolism studies with special emphasis on the recent advancement in the field.