Propolis in the management of cardiovascular disease.

BACKGROUND: Propolis is a resinous compound that is obtained from honey bees. It consists of numerous chemical constituents that impart different therapeutic action. The heart is the core of the body and cardiovascular disease (CVD) is a burden for the human being. This article emphasizes how propolis is fruitful in the management of various CVDs. SCOPE AND APPROACH: This review focuses on how various constituents of the propolis (such as terpenes, flavonoids, phenolics, etc.) impart cardio protective actions. KEY FINDING AND CONCLUSION: With the support of various clinical trials and research outcomes, it was concluded that propolis owns niche cardio protective properties that can be a boon for various cardiac problems (both in preventive and therapeutic action) such as atherosclerosis, excessive angiogenesis, hypertension, and many more.

Combatting infectious diarrhea: innovations in treatment and vaccination strategies.

INTRODUCTION: The escalating prevalence of infectious diseases is an important cause of concern in society. Particularly in several developing countries, infectious diarrhea poses a major problem, with a high fatality rate, especially among young children. The condition is divided into four classes, namely, acute diarrhea, invasive diarrhea, acute bloody diarrhea, and chronic diarrhea. Various pathogenic agents, such as bacteria, viruses, protozoans, and helminths, contribute to the onset of this condition. AREAS COVERED: The review discusses the scenario of infectious diarrhea, the prevalent types, as well as approaches to management including preventive, therapeutic, and vaccination strategies. The vaccination techniques are extensively discussed including the available vaccines, their advantages as well as limitations. EXPERT OPINION: There are several approaches available to develop new-improved vaccines. In addition, route of immunization is important and aerosols/nasal sprays, oral route, skin patches, powders, and liquid jets to minimize needles can be used. Plant-based vaccines, such as rice, might save packing and refrigeration costs by being long-lasting, non-refrigerable, and immunogenic. Future research should utilize predetermined PCR testing intervals and symptom monitoring to identify persistent pathogens after therapy and symptom remission.

Microalgae-Based Disinfectant Formulation for Aseptic Processing of Ethiopian Ingredient-Sourced Functional Bread and Its Molecular Docking Analysis to Reduce Hypernatremia.

The global prevalence of food-borne infections has become a major concern. Food-borne pathogens like Campylobacter jejuni, Salmonella enterica, and Clostridium botulinum cause food poisoning and even mortality, necessitating the maintenance of aseptic conditions during food processing. The sterilization of food processing facilities often requires chemical and heat treatment. The formulation of many chemical-based disinfectants includes chemicals generating toxic and carcinogenic by-products. The microalgae like Chlorella spp. reportedly exhibit antimicrobial activity and therefore, can be used for formulating safer and eco-friendly natural sanitizers. This study aims to aseptically prepare functional bread using Ethiopian ingredients, highlighting the application of microalgae-based disinfectant formulation and various disinfection techniques. The functional bread was designed to be potentially effective in reducing hypernatremia condition which is indicative of high levels of sodium in serum that can cause an array of symptoms including deaths in serious cases. The physico-chemical and sensory properties of the designed functional bread were analyzed. The interaction of phytochemicals in the ingredients with the target receptor (Vasopressin V2 receptor) and their drug-likeness were determined using molecular docking and Lipinski's rule of five analyses. The results suggest that the designed functional bread incorporating Ethiopian ingredients may serve as an effective dietary strategy to prevent hypernatremia. Aseptic processing of the bread ensures longer shelf life and prevention of spoilage by food pathogens.

Toxicity of chemical-based hand sanitizers on children and the development of natural alternatives: a computational approach.

The unintended exposure of children to hand sanitizers poses a high risk of potentially fatal complications. Skin irritation, dryness, cracking, peeling, hypoglycemia, apnea, and acidosis are examples of unintended consequences of hand sanitizer. The sanitizer reportedly kills normal microbial flora on hands, which usually promotes innate immunity among children under 12. Children are more susceptible to the toxicity associated with the chemical constituents of marketed chemical-based hand sanitizers; however, the studies to develop sanitizer formulations for children are rudimentary. The adverse events limit the use of hand sanitizers specifically in children because of their sensitive and delicate skin. Additionally, it is reported that many chemical-based hand sanitizer formulations, especially alcohol-based ones may also contain contaminants like methanol, acetaldehyde, benzene, isopropanol, and ethyl-acetate. These contaminants are found to be hazardous to human health exhibiting toxicity on ingestion, inhalation, or dermal exposure, especially in children. Therefore, it is important to design novel, innovative, safer sanitizer formulations for children. The study aims to discuss the toxic contaminants in chemical-based sanitizer formulations and propose a design for novel herbal formulations with minimal toxicity and adverse effects, especially for children. The review focuses on ADMET analysis of the common contaminants in hand sanitizers, molecular docking, Lipinski's rule of five analysis, and molecular simulation studies to analyze the efficacy of interaction with the receptor leading to anti-microbial activity and drug-likeness of the compound. The in silico methods can effectively validate the potential efficacy of novel formulations of hand sanitizers designed for children as an efficient alternative to chemical-based sanitizers with greater efficacy and the absence of toxic contaminants.

The utilization of natural eco-benign sources for sustainable management to preserve hides and docking analysis of identified potential phytochemicals.

The industrial leather sector is one of the most ancient industries globally and continues to influence the global economic system in contemporary times significantly. Regardless of income, the leather sector is widely recognized as a significant environmentally detrimental sector because of the utilization of materials involved in preserving and processing leather. Raw hides, the primary ingredient in the leather industry, are highly susceptible to microbial attack once they have been flayed from animals. The current review provides information about the diverse operational processes employed in the beam house to produce leather goods. This paper primarily focuses on the preservation of various types of hides, including those derived from goats, cows, sheeps, deers, pigs, and other species, with particular emphasis on bio-based preservation methods. It also discusses various salt and salt-free curing methods along with their limitations. Furthermore, it reviews a viable and economically advantageous option for preserving animal hides using plant and microbial sources. Moreover, it focuses on investigating the molecular docking interactions between three critical enzymes in the phytochemical synthesis pathway, namely phenylalanine ammonia-lyase, phosphomevalonate kinase, and dihydroflavonol 4-reductase, and a set of specific modulators, followed by ADMET analyses. The objective was to determine the optimal binding affinity score for these enzyme-modulator complexes through virtual screening. The depiction of protein-ligand interactions offers potential benefits for future research endeavours, as well as valuable insights into the identification of modulators and the evaluation of the potency and efficacy of phytochemicals in the preservation of hides.

Recent Review on the Extraction and Qualitative Assay of Cysteine and Other Amino Acids from Vellore Feather Waste and Molecular Docking Studies of Cysteine for Pharmacological Applications.

Products produced from waste are a relatively recent innovation. Feather substrates are abundant in keratin content and improper disposal can cause ecosystem contamination. However, these pollutants can be transformed into value-added products for industrial application. Physical, chemical and cutting-edge microbiological methods were utilized for decomposing keratin and aid in the identification and estimation of amino acids from poultry feather wastes. These beneficial approaches are receiving more attention due to their retrieval of harmless and value added byproducts. These keratin-based compounds are used widely in pharmaceutical, livestock feed, fertilizer, and a variety of other industrial sectors. Since keratin is primarily consisting of amino acids, it can be utilized to affirm and estimate the amino acids in these feather substrates. This study primarily highlights the various methodologies employed for the qualitative estimation of amino acids in feather waste samples and the inhibitory activity of keratinase enzyme by EDTA and pepstatin in order to accumulate amino acids for drug delivery purpose and their importance in various pharmaceutical industries. In addition to that, molecular docking studies of cysteine with many standard pharmaceutical drugs like acetaminophen, pethidine, methylphenidate, carbamazepine, cillin and amlodipine were performed using autodock to demonstrate how cysteine greatly reduces conventional drug toxicity and its side effects.

In Silico Study of Enzymatic Degradation of Bioplastic by Microalgae: An Outlook on Microplastic Environmental Impact Assessment, Challenges, and Opportunities.

Microplastics are tiny pieces of non-biodegradable plastic that can take thousands of years to break down. As microplastics degrade, they release harmful compounds into the environment, which can be found in the surroundings. The microplastics found in the environment are hard to detect and remove because of their small particle sizes. Microplastics cannot decompose naturally, so they accumulate in the environment and cause pollution. As a result, bioplastics can be produced from a vast array of substrates, including biopolymers, citrus peels, leather, and feather wastes. Blue-green microalgae namely Arthrospira platensis (spirulina) contains enzymes such as laccase and catalase which can be responsible for the degradation of bioplastics. In our study, we performed molecular docking to identify the binding affinities of different enzymes such as laccase and catalase with different substrates, focusing on determining the most suitable substrate for enhancing enzyme activity for degradation of bioplastics. The analysis revealed that veratryl alcohol is the most suitable substrate for laccase, whereas lignin is the more preferred substrate for catalase with the highest binding affinity score of - 5.9 and - 8.1 kcal/mol. Moreover, degradation, challenges, opportunities, and applications of bioplastics in numerous domains such as cosmetics, electronics, agriculture, medical, textiles, and food industries have also been highlighted.

The sustainable conversion of floral waste into natural snake repellent and docking studies for antiophidic activity.

Snakes play an important role as predators, prey, ecosystem regulators and in advancing the human economy and pharmaceutical industries by producing venom-based medications such as anti-serums and anti-venoms. On the other hand, snakebites are responsible for over 120,000 annual fatalities; due to snakebites people lose their lives and suffer from diseases such as snake envenoming, epilepsy, and symptoms such as punctures, swelling, haemorrhage, bruising, blistering, and inflammation. Moreover, there are several challenges associated with different interventions for managing snakebites. Therefore, finding a natural way of repelling snakes without harming them will save lives and decrease the disease's symptoms. Usually, snakes are exacerbated by noxious odours and shrill sounds. There are various strategies to repel snakes, including chemical, natural, and electronic repellents being the most prevalent. Chemical snake repellents such as mothballs, sulphur powder, and cayenne pepper act as a barrier; natural snake repellents produce a pungent and foul smell, while electronic repellents generate high-frequency ultrasonic waves to repel snakes. On the other hand, anti-serums are available commercially to prevent the adverse effects of snakebite, which are species-specific, expensive, have inadequate pharmacology and impaired interaction with the immune system. Similarly, there are monovalent or polyvalent anti-serums used for the production of anti-venom depending on the snake species and the number of snakebites occurred in that area, e.g., Soro antibotropicocrotalico contains specific antibodies for Pit vipers and rattlesnakes, and Antielapidico targets coral snakes. The purpose of this review is to investigate natural, effective, and inexpensive snake-repellent from Vellore Institute of Technology (VIT) floral waste, which can be mixed with natural products such as vinegar, citronella, cinnamon, garlic, cedar, and clove and allowed for bacterial degradation which will lead to the release of several gases during floral waste degradation, including ammonia, sulphur, manganese, selenium, and gallic acid due to bacterial growth like Proteus, Bacillus, Streptococcus, etc. We assumed to convert these gases into liquid form using Linde's technique which may repel snakes. Further, molecular docking studies were performed on snake venom toxins (Phospholipase A2 (PDB-1MG6), Protein Cytotoxin II (PDB-1CB9), α-Dendrotoxins (PDB-1DTX), Neurotoxin from cobra venom (PDB-1CTX) and Cardiotoxin III (PDB-2CRS). Phytocompounds of Vellore degraded floral waste from GC-MS analysis (Tetracosane, 12, Oleanen-3-yl Acetate, (3-Alpha), Eicosane-7-Hexyl, Octadecane,3-Ethyl-5(2-Ethyl Butyl), Nonadecane,4-Methyl, Hexatriacontane and Nonacosane) were used as a ligand to determine their binding affinity with venom proteins and may be assumed to be used as an antidote for snakebite. Finally, we analysed that 12-oleanen-3yl acetate,3-α (CID-45044112) a triterpenoid showing a maximum binding affinity with all snake venom proteins (-13.8k/cal) with Phospholipase A2 (PLA2), Cardiotoxin-II (-8.2k/cal), Dendrotoxin (-12.1 k/cal), Cardiotoxin-III (-8.2 kcal/mol) and alpha-Neurotoxin (-11.0 kcal/mol), which may have potential to counteract the adverse effects caused by snakebites, however, in-vitro and in-vivo studies still challenging tasks for our further analysis. Overall, we propose an innovative method for the sustainable conversion of floral waste into snake repellent, as well as molecular docking studies were performed with phytocompounds and snake venom proteins for antiophidic activity, which can be experimentally investigated further to confirm its use as anti-venom for snakebites.

The Emergence of Hybrid Variants of SARS-CoV-2: Towards Hybrid Immunity.

In this review work, the authors emphasize the discussion on different emerging variants of SARS-CoV-2 and vaccine effectiveness against them [...].

Post-COVID-19 Fungal Infection in the Aged Population.

Coronavirus disease 2019 (COVID-19) infection is currently a great cause of concern for the healthcare sector around the globe. SARS-CoV-2 is an RNA virus that causes a serious infection that is associated with numerous adverse effects and multiple complications associated with different organs and systems during its pathogenic cycle in humans. Individuals affected by COVID-19, especially elderly populations and immunocompromised people, are greatly vulnerable to opportunistic fungal pathogens. Aspergillosis, invasive candidiasis, and mucormycosis are widespread fungal coinfections in COVID-19 patients. Other fungal infections that are rare but are exhibiting increased incidence in the current scenario include infections caused by Pneumocystis jirovecii, Histoplasma sp., Cryptococcus sp., etc. By producing virulent spores, these pathogens increase the severity of the disease and increase the morbidity and fatality rates in COVID-19 patients globally. These infections generally occur in patients recovering from COVID-19 infection, resulting in rehospitalization. Older and immunocompromised individuals are at higher risk of developing opportunistic fungal infections. This review focuses on understanding the opportunistic fungal infections prevalent in COVID-19 patients, especially elderly people. We have also highlighted the important preventive methods, diagnostic approaches, and prophylactic measures for fungal infections.

Computational Modelling and Molecular Docking of Industrial Leather Enzymes.

Leather is one of the widely traded commodities globally. It is a strategically important sector for the economic and industrial development of the country. However, the leather industry is perceived as a highly polluting industry. It produces huge amounts of solid and liquid wastes, and if these wastes are not properly treated and disposed of, then it tends to deteriorate the quality of soil and water, as well as cause emanations of smell and noxious gases into the surrounding. The current paper provides information about industrial leather enzymes, primarily collagenase, tannase, and lecithinase. In this study, enzymes such as collagenase, tannase, and lecithinase had a pivotal role in leather industries and their action in the bioremediation of leather effluents was further analysed and docked with a diverse range of compounds (ligands), with an optimal binding affinity score was determined. All interactions between protein ligands were depicted, which will help us with future research. Furthermore, this method can be tested practically, and other parameters can be studied in the future. Further, applications of enzymes and their hydrolyse by-products have also been highlighted in a variety of industries, including the pharmaceutical, cosmetic, agricultural, medical, and food sectors. Subsequently, this finding provides an innovative and broader goal for various sectors in terms of sustainability, stabilisation, and identifying research gaps that can guide modern industries and research scientists.

Recent review of COVID-19 management: diagnosis, treatment and vaccination.

The idiopathic Coronavirus disease 2019 (COVID-19) pandemic outbreak caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has reached global proportions; the World Health Organization (WHO) declared it as a public health emergency during the month of January 30, 2020. The major causes of the rise of new variants of SARS-CoV-2 are genetic mutations and recombination. Some of the variants with high infection and transmission rates are termed as variants of concern (VOCs) like currently Omicron variants. Pregnant women, aged people, and immunosuppressed and compromised patients constitute the most susceptible human population to the SARS-CoV-2 infection, especially to the new evolving VOCs. To effectively manage the pathological condition of infection, the focus should be directed towards prevention and prophylactic approach. In this narrative review, we aimed to analyze the current scenario of COVID-19 management and discuss the treatment and prevention strategies. We also focused on the complications prevalent during the COVID-19 and post-COVID period and to discuss the novel approaches developed for mitigation of the global pandemic. We have also emphasized on the COVID-19 management approaches for the special population including children, pregnant women, aged groups, and immunocompromised patients. We conclude that the advancements in therapeutic and pharmacological domains have provided opportunities to develop and design novel diagnosis, treatment, and prevention strategies. New advanced techniques such as RT-LAMP, RT-qPCR, High-Resolution Computed Tomography, etc., efficiently diagnose patients with SARS-CoV-2 infection. In the case of treatment options, new drugs like paxlovid, combinations of ß-lactum drugs and molnupiravir are found to be effective against even the new emerging variants. In addition, vaccination is an essential approach to prevent the infection or to reduce its severity. Vaccines for against COVID-19 from Comirnaty by Pfizer-BioNTech, SpikeVax by Moderna, and Vaxzevria by Oxford-AstraZeneca are approved and used widely. Similarly, numerous vaccines have been developed with different percentages of effectiveness against VOCs. New developments like nanotechnology and AI can be beneficial in providing an efficient and reliable solution for the suppression of SARS-CoV-2. Public health concerns can be efficiently treated by a unified scientific approach, public engagement, and better diagnosis.

Teff Grain-Based Functional Food for Prevention of Osteoporosis: Sensory Evaluation and Molecular Docking Approach.

Osteoporosis is a highly prevalent skeletal disorder, which is characterized by compromised bone strength predisposing to an increased risk of fracture. The medical approaches presently followed for the prevention and treatment are associated with several side effects. Thus, it becomes important to design alternatives that are safer, economical, and easy to use. Plants provide us with a beneficial and effective option for such designs. They are rich in nutrients and phytochemicals that can be used to target signalling pathways to prevent the development of disease. For this purpose, we used Ethiopian grain, teff as a preventive strategy for osteoporosis as it has a favourable nutritional profile. In our study, we focussed on the preparation of functional multi-grain flour with Ethiopian grain, teff, and other ingredients that have a positive effect on bone health. A cookie was prepared from the multi mix flour and to analyze its shelf life and acceptance we performed a sensory evaluation where we observed no significant difference in the cookies' characteristics during the period of study. We also performed Molecular Docking of eight flavonoid polyphenols selected from the nutritional profile of the ingredients to understand their binding affinity, interaction with the target, receptor activator of nuclear factor kappa-Β ligand (RANKL), and prophylactic or therapeutic effects in the prevention of osteoporosis. It was found that all 8 flavonoid polyphenols bound with RANKL, at least at one of the crucial binding sites and so can be used for the prevention of osteoporosis.

Immunological Studies to Understand Hybrid/Recombinant Variants of SARS-CoV-2.

The zoonotic SARS-CoV-2 virus was present before the onset of the pandemic. It undergoes evolution, adaptation, and selection to develop variants that gain high transmission rates and virulence, resulting in the pandemic. Structurally, the spike protein of the virus is required for binding to ACE2 receptors of the host cells. The gene coding for the spike is known to have a high propensity of mutations, as a result generating numerous variants. The variants can be generated by random point mutations or recombination during replication. However, SARS-CoV-2 can also produce hybrid variants on co-infection of the host by two distinct lineages of the virus. The genomic sequences of the two variants undergo recombination to produce the hybrid variants. Additionally, these sub-variants also contain numerous mutations from both the parent variants, as well as some novel mutations unique to the hybrids. The hybrid variants (XD, XE, and XF) can be identified through numerous techniques, such as peak PCR, NAAT, and hybrid capture SARS-CoV-2 NGS (next generation sequencing) assay, etc., but the most accurate approach is genome sequencing. There are numerous immunological diagnostic assays, such as ELISA, chemiluminescence immunoassay, flow-cytometry-based approaches, electrochemiluminescence immunoassays, neutralization assays, etc., that are also designed and developed to provide an understanding of the hybrid variants, their pathogenesis, and other reactions. The objective of our study is to comprehensively analyze the variants of SARS-CoV-2, especially the hybrid variants. We have also discussed the techniques available for the identification of hybrids, as well as the immunological assays and studies for analyzing the hybrid variants.

Characterisation of pectin and optimization of pectinase enzyme from novel Streptomyces fumigatiscleroticus VIT-SP4 for drug delivery and concrete crack-healing applications: An eco-friendly approach.

Pectinases are enzymes which are widely distributed in microbes that are present in pectin enriched sites. The agro-industrial residues can be utilized in the industrial scale for low-cost and efficient pectinase production in an eco-friendly approach. This study employs low-cost substrates (i.e. culinary fruit peels) for maximum pectinase production from novel Streptomyces fumigatiscleroticus VIT-SP4. The extraction and characterization of pectin from different fruit peels were investigated and pectinase activity was analyzed. The orange pectin gave maximum pectinase activity of about 45.93 (U/mL). Further, statistical optimization of process parameters was studied by using Taguchi method showed optimum values of pH-6, temperature -35 °C, orange pectin% - 2.5, incubation time- 48 h and RPM- 200 rpm and pectinase activity was found to be 98.65 (U/mL). The response surface methodology (RSM) was used for the optimization of media components which revealed that starch -1.17%, yeast extract-2%, and orange pectin% - 0.75% produces maximum pectinase of about 170.05 (U/mL). The drug-delivery study showed drug release was not observed at initial pH 3 after 4 h. The immediate drug release was noted at pH 6 caused due to disintegration of pectin by the pectinase activity. The self-healing of cracks by spray culture technique was investigated. The crack healing was observed up to 0.50 mm wide after 12 days. This confirms the ability of actinomycete spores to survive and they react to form calcite complex directly helps in crack healing process. This low-cost microbial pectinase can be used in drug delivery and concrete crack-healing applications sectors in future.

Taguchi analysis and asymmetric keto-reduction of acetophenone and its derivatives by soil filamentous fungal isolate: Penicillium rubens VIT SS1.

Microbial asymmetric reduction of ketone is an efficient tool for the synthesis of chiral alcohols. This research focuses on exploring the soil fungal isolates for their ability toward the keto reduction of acetophenone and its derivatives to their corresponding chiral alcohols using growing cells. Bioreduction of acetophenone, 4-fluoro acetophenone, 4-methyl acetophenone, and 3-hydroxy acetophenone was carried out using different fungal cultures isolated from soil. Among the fungal isolates, Penicillium sp. and Aspergillus sp. showed significant bioconversion with varying enantio-selectivity. However, the Penicillium sp. has shown the maximum ability of bioreduction. The potential isolate was characterized using the internal transcribed spacer (ITS) region and found to be Penicillium rubens VIT SS1 (Genbank accession number: MK063869.1), which showed higher conversion and selectivity > 90%. The biocatalyst production and the reaction conditions were optimized using Taguchi analysis. The process conditions such as pH, temperature, media components, cosolvent, and substrate dosing were evaluated for the bioreduction of 3-hydroxy acetophenone, which is a key chiral intermediate of Phenylephrine and Rivastigmine using P. rubens VIT SS1. This study concludes about the potential of fungal cultures for sustainable synthesis of key chiral intermediates of Phenylephrine and Rivastigmine, similarly many aromatic chiral alcohols in simpler, novel, and cost-effective manner.

In vitro drug metabolism and pharmacokinetics of a novel thiazolidinedione derivative, a potential anticancer compound.

Thiazolidinediones are known for their activity against Type 2 diabetes and are currently being repurposed for their potent anti-cancer activity. In the present study, we have assessed in vitro metabolic properties and in vivo pharmacokinetic parameters of a novel thiazolidinedione derivative, BIT-15-67, a potential anticancer compound. BIT-15-67 showed low solubility in aqueous buffers at different pH values. The permeability was determined across the Caco-2 monolayer and BIT-15-67 showed high permeability and an efflux ratio of less than 2 suggesting that it is not a substrate of the efflux transporters (P-gp & BCRP). The plasma protein binding was evaluated by equilibrium dialysis and the compound exhibited moderate binding to mouse and rat plasma proteins. BIT-15-67 was stable (half-life > 30 min.) in mouse, rat, dog and human liver microsomes and unstable (half-life <15 min.) in rat hepatocytes suggesting possible Phase II metabolism. Liquid chromatography-tandem mass spectrometry was used to identify Phase I and Phase II metabolites. One of each Phase I and Phase II metabolites have been identified in rat hepatocytes samples. The BIT-15-67 is not an inhibitor of CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4. The PK parameters were determined in both male and female Wistar rats after single intravenous dose administration of BIT-15-67. In rats, the mean plasma clearance of BIT-15-67 was higher in males than in females and the terminal plasma elimination half-life was shorter in males than in females. The compound was highly distributed in the tissues. Overall, the absolute oral bioavailability was 5-fold higher in females (38 %) than in males (7 %). In female nude mice with tumors, BIT-15-67 was well distributed among the collected tissues with the highest concentration in the liver. The ratio of the concentrations in tumor vs. the plasma was 0.5 which could be an important attribute in the development of the compound for anti-cancer research.

Validation of a sensitive simultaneous LC-MS/MS method for the quantification of novel anti-cancer thiazolidinedione and quinazolin-4-one derivatives in rat plasma and its application in a rat pharmacokinetic study.

Thiazolidinediones and quinazolin-4-ones compounds, previously known for their activity against Type 2 diabetes and antifungal activity respectively, are currently being investigated for their anti-cancer activity. The determination of pharmacokinetic parameters for these two classes of compounds using a simultaneous chromatographic method with a low detection limit is a challenge. In this study, a highly sensitive and simultaneous LC-MS/MS-based bioanalytical method was developed and validated in rat plasma for the estimation of four novel anti-cancer compounds, BIT-15-67 and BNT-11, belonging to the Thiazolidinedione class, and BNUA-108 and BNUA-48, from the quinazolin-4-one class. The analytes were extracted from plasma samples by protein precipitation and separated on a short reverse phase Hypersil Phenyl BDS, 50 × 4.6 mm, 2.4 µm column at a column oven temperature of 40 °C. An isocratic mobile phase, a 20:80 (v/v) mixture of 5 mM ammonium acetate solution and acetonitrile containing 0.1% formic acid, was used for the elution at a flow rate of 0.4 mL/min. The analytes and internal standard, sulfaphenazole, were quantified in the multiple reaction monitoring mode using positive electrospray ionization with specific pair of mass by charge ratio. All standard validation parameters were assessed as per current bioanalytical method validation guidelines in rat plasma. The area response for the four analytes was found to be linear over the concentration range of 1.00 to 1000 ng/mL in rat plasma. The signal to noise at LLOQ of 1 ng/mL was adequate for application to different pre-clinical studies. The intra- and inter-day precision were <11% and accuracy deviated -1.8 to 9.60% from the nominal. The mean recovery was high (about 90%) and consistent for all the analytes over the linear dynamic range of the method. This simple, robust and validated method can be employed to determine the rat plasma concentrations of the four selected anticancer compounds in preclinical studies such as the pharmacodynamic and the pharmacokinetic studies including tissue distribution and excretion, and the toxicokinetic studies. In this study, pharmacokinetic parameters were determined using this method for all the four compounds individually following intravenous administration in rats.