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Continuous-flow chemistry has become a mainstream process and a notable trend among emerging technologies for drug synthesis. It is routinely used in academic and industrial laboratories to generate a wide variety of molecules and building blocks. The advantages it provides, in terms of safety, speed, cost efficiency and small-equipment footprint compared to analog batch processes, have been known for some time. What has become even more important in recent years is its compliance with the quality objectives that are required by drug-development protocols that integrate inline analysis and purification tools. There can be no doubt that worldwide government agencies have strongly encouraged the study and implementation of this innovative, sustainable and environmentally friendly technology. In this brief review, we list and evaluate the development and applications of continuous-flow processes for antibiotic synthesis. This work spans the period of 2012-2022 and highlights the main cases in which either active ingredients or their intermediates were produced under continuous flow. We hope that this manuscript will provide an overview of the field and a starting point for a deeper understanding of the impact of flow chemistry on the broad panorama of antibiotic synthesis.

Human liver subcellular fractions, including liver microsomes (HLM), liver cytosol fractions, and S9 fractions, are extensively utilized in in vitro assays to predict liver metabolism. The S9 fractions are supernatants of human liver homogenates that contain both microsomes and cytosol, which include most cytochrome P450 (CYP) enzymes and soluble phase II enzymes such as glucuronosyltransferases and sulfotransferases. This study reports on the direct electrochemistry and biocatalytic features of redox-active enzymes in S9 fractions for the first time. We investigated the electrochemical properties of S9 films by immobilizing them onto a high-purity graphite (HPG) electrode and performing cyclic voltammetry under anaerobic (Ar-saturated) and aerobic (O2-saturated) conditions. The heterogeneous electron transfer rate between the S9 film and the HPG electrode was found to be 14 ± 3 s−1, with a formal potential of −0.451 V vs. Ag/AgCl reference electrode, which confirmed the electrochemical activation of the FAD/FMN cofactor containing CYP450-reductase (CPR) as the electron receiver from the electrode. The S9 films have also demonstrated catalytic oxygen reduction under aerobic conditions, identical to HLM films attached to similar electrodes. Additionally, we investigated CYP activity in the S9 biofilm for phase I metabolism using diclofenac hydroxylation as a probe reaction and identified metabolic products using liquid chromatography–mass spectrometry (LC-MS). Investigating the feasibility of utilizing liver S9 fractions in such electrochemical assays offers significant advantages for pharmacological and toxicological evaluations of new drugs in development while providing valuable insights for the development of efficient biosensor and bioreactor platforms. [ABSTRACT FROM AUTHOR]

Skeleton editing has rapidly advanced as a synthetic methodology in recent years, significantly streamlining the synthesis process and gaining widespread acceptance in drug synthesis and development. This field encompasses diverse ring reactions, many of which exhibit immense potential in skeleton editing, facilitating the generation of novel ring skeletons. Notably, reactions that involve the cleavage of two distinct rings followed by the reformation of new rings through ring insertion play a pivotal role in the construction of novel ring skeletons. This article aims to compile and systematize this category of reactions, emphasizing the two primary reaction types and offering a thorough exploration of their associated complexities and challenges. Our endeavor is to furnish readers with comprehensive reaction strategies, igniting research interest and injecting fresh impetus into the advancement of this domain. [ABSTRACT FROM AUTHOR]

Aim/Background: The Cytochrome P450 (CYP) enzyme family, consisting of 57 distinct genes, plays a pivotal role in various biological processes. These genes encode enzymes with multifaceted functions, impacting vital processes like arachidonic acid metabolism, xenobiotic detoxification, eicosanoids production, and drug metabolism. Moreover, CYP enzymes are indispensable for the synthesis of bile acids, steroids, and numerous metabolic pathways. They also participate in the hydroxylation of retinoic acid, illustrating their remarkable versatility. While some CYP enzymes still have undiscovered functions, their role continues to captivate researchers across diverse domains. Additionally, mutations in CYP genes can give rise to inborn metabolic disorders, leading to clinically significant diseases, underscoring the importance of CYP enzymes in maintaining metabolic equilibrium and overall well-being. Beyond their initial association with hepatic drug detoxification, recent research has unveiled a broader spectrum of enzymatic processes undertaken by cytochrome P450. The complexity of CYP enzymes is progressively unfolding, emphasizing their clinical significance and opening new avenues for drug development, precision medicine, and patient care. methodology: This abstract presents a comprehensive overview of the literature on cytochrome P450 enzymes and their diverse functions. The information was gathered from a wide range of sources, including scientific articles, textbooks, and research publications. The analysis covers the genetic basis of CYP enzymes, their roles in various metabolic pathways, and the clinical implications of CYP gene mutations. Additionally, the abstract highlights the recent advances in our understanding of CYP enzymes and their expanding role in maintaining human life. Results: The results of this study showcase the remarkable diversity of functions performed by cytochrome P450 enzymes. They are intricately involved in processes critical to human health, ranging from drug metabolism to the synthesis of vital biomolecules. The study underlines the clinical significance of CYP enzymes, as mutations in these genes can lead to severe metabolic disorders and diseases. Furthermore, it emphasizes the evolving understanding of CYP enzyme complexity and its implications for drug development, precision medicine, and patient care. Conclusion: In conclusion, this abstract sheds light on the exceptional versatility and clinical importance of cytochrome P450 enzymes. Their diverse functions have far-reaching consequences for human health, making them a focal point of research in pharmacology and medicine. The evolving understanding of CYP enzyme complexity paves the way for future advancements in drug development and personalized patient care, offering new possibilities for enhancing the well-being of individuals and populations. This research underscores the significance of CYP enzymes and their potential to revolutionize the fields of medicine and pharmacology. [ABSTRACT FROM AUTHOR]

Imidazo[1,2-a]pyrimidine derivatives are a class of biologically active parent nucleus structures containing both imidazole and pyrimidine rings, which are important intermediates for the synthesis of a variety of drugs. This class of structures has attracted a lot of attention of researchers, and more and more synthetic strategies have been developed and reported. Currently, the main synthetic strategies are one-pot multi-component methods, microwave radiation, ultrasonic irradiation and other auxiliary Knoevenagel condensation and Michael addition methods to obtain these compounds, with the aim of achieving high yields, low time consumption and environmental friendliness. This review comprehensively summarizes the various synthetic methods as well as mechanism recently reported for the construction of this remarkable backbone structure and further used for the development of new chemical synthesis strategies and drug development as it has a wide range of applications in medicinal chemistry. In order to provide some impetus to the research of synthetic methods for this structure. [ABSTRACT FROM AUTHOR]

Automating retrosynthesis with artificial intelligence expedites organic chemistry research in digital laboratories. However, most existing deep-learning approaches are hard to explain, like a "black box" with few insights. Here, we propose RetroExplainer, formulizing the retrosynthesis task into a molecular assembly process, containing several retrosynthetic actions guided by deep learning. To guarantee a robust performance of our model, we propose three units: a multi-sense and multi-scale Graph Transformer, structure-aware contrastive learning, and dynamic adaptive multi-task learning. The results on 12 large-scale benchmark datasets demonstrate the effectiveness of RetroExplainer, which outperforms the state-of-the-art single-step retrosynthesis approaches. In addition, the molecular assembly process renders our model with good interpretability, allowing for transparent decision-making and quantitative attribution. When extended to multi-step retrosynthesis planning, RetroExplainer has identified 101 pathways, in which 86.9% of the single reactions correspond to those already reported in the literature. As a result, RetroExplainer is expected to offer valuable insights for reliable, high-throughput, and high-quality organic synthesis in drug development. Automating retrosynthesis prediction in organic chemistry is a major application of ML. Here the authors present RetroExplainer, which offers a high-performance, transparent and interpretable deep-learning framework providing valuable insights for drug development. [ABSTRACT FROM AUTHOR]

Molecular editing, including peripheral and skeletal editing, is an area of interest in the field of organic synthesis. Typically, the peripheral editing of C―H bonds is conventionally used for molecular editing. By contrast, skeletal editing is a novel method, which has broader applications in total synthesis and drug development, with advantages of easy operation, high specificity, and simple synthetical routes. This review introduces two skeletal editing examples using common reactions. By comparisons with conventional methods, the applications of molecular editing in drug synthesis are revealed. Moreover, this review predicts the future applications of molecular editing by presenting some state-of-the-art developments in this field. [ABSTRACT FROM AUTHOR]

The processes used by academic and industrial scientists to discover new drugs have recently experienced a true renaissance, with many new and exciting techniques being developed over the past 5–10 years alone. Drug design and discovery, and the search for new safe and well-tolerated compounds, as well as the ineffectiveness of existing therapies, and society's insufficient knowledge concerning the prophylactics and pharmacotherapy of the most common diseases today, comprise a serious challenge. This can influence not only the quality of human life, but also the health of whole societies, which became evident during the COVID-19 pandemic. In general, the process of drug development consists of three main stages: drug discovery, preclinical development using cell-based and animal models/tests, clinical trials on humans and, finally, forward moving toward the step of obtaining regulatory approval, in order to market the potential drug. In this review, we will attempt to outline the first three most important consecutive phases in drug design and development, based on the experience of three cooperating and complementary academic centers of the Visegrád group; i.e., Medical University of Lublin, Poland, Masaryk University of Brno, Czech Republic, and Comenius University Bratislava, Slovak Republic. [ABSTRACT FROM AUTHOR]

Droplet-based microfluidics offer great opportunities for applications in various fields, such as diagnostics, food sciences, and drug discovery. A droplet provides an isolated environment for performing a single reaction within a microscale-volume sample, allowing for a fast reaction with a high sensitivity, high throughput, and low risk of cross-contamination. Owing to several remarkable features, droplet-based microfluidic techniques have been intensively studied. In this review, we discuss the impact of droplet microfluidics, particularly focusing on drug screening and development. In addition, we surveyed various methods of device fabrication and droplet generation/manipulation. We further highlight some promising studies covering drug synthesis and delivery that were updated within the last 5 years. This review provides researchers with a quick guide that includes the most up-to-date and relevant information on the latest scientific findings on the development of droplet-based microfluidics in the pharmaceutical field. [ABSTRACT FROM AUTHOR]

The Open Research Biopharmaceutical Internships Support project (ORBIS) was a response to the scientific, economic, and social challenge of increasing the effectiveness and productivity of the drug development process, both for innovative and (super)generic drugs. The overarching objective of the ORBIS project was to form a transnational and intersectoral cooperation network of academic and industrial organisations delivering a joint research programme. The research aimed at improving the preclinical pathway of drug development and manufacturing, focusing on technological and methodological improvements of the existing processes. The participating staff from all institutions have developed new skills, were exposed to new work and research environments, and have significantly broadened their career perspectives. More than 450 months of secondments were completed, and over 175 early-stage and experienced researchers participated in the exchange. This review aims to present some aspects of the scientific, training, and organisational activities of the consortium, bringing together representatives of both the academic sector as well as small and medium-sized pharmaceutical enterprises. [ABSTRACT FROM AUTHOR]

A new series of heterofused thiazolo/pyrazinedione hybrids were designed, synthetized and their cytotoxicity potential was evaluated against different cancer cell lines. Besides, drug‐likeness and in silico ADME/Tox studies were carried out using free server tools. The heterofused hybrids 3‐aryl‐thiazolo[3,4‐a]pyrazine‐5,8‐diones 3 a–o were obtained in moderate to good yields (30–80 %) and with high diastereomeric ratio. Notably, hybrids 3 g–j showed good cytotoxic effect on Hep‐G2 cancer cells IC50 (28.6±4.9 μM, 54.2±15.4 μM, 95.2±1.9 μM, 68.6±1.6 μM) respectively, while compounds 3 l–m were cytotoxic on B16F10 melanoma cancer cells IC50 (20.9±5.3 μM, 30.0±6.4 μM). Furthermore, the new compounds were relatively non‐toxic on Vero cells. In silico drug‐likeness and ADME/Tox studies suggest optimal pharmacokinetic profile for all hybrids. Altogether, these new heterofused hybrids could be considered as promising scaffolds in cancer chemotherapy. [ABSTRACT FROM AUTHOR]

While there are many clinical drugs for prophylaxis and treatment, the search for those with low or no risk of side effects for the control of infectious and non-infectious diseases is a dilemma that cannot be solved by today's traditional drug development strategies. The need for new drug development strategies is becoming increasingly important, and the development of new drugs from traditional medicines is the most promising strategy. Many valuable clinical drugs have been developed based on traditional medicine, including drugs with single active ingredients similar to modern drugs and those developed from improved formulations of traditional drugs. However, the problems of traditional isolation and purification and drug screening methods should be addressed for successful drug development from traditional medicine. Advances in microfluidics have not only contributed significantly to classical drug development but have also solved many of the thorny problems of new strategies for developing new drugs from traditional drugs. In this review, we provide an overview of advanced microfluidics and its applications in drug development (drug compound synthesis, drug screening, drug delivery, and drug carrier fabrication) with a focus on its applications in conventional medicine, including the separation and purification of target components in complex samples and screening of active ingredients of conventional drugs. We hope that our review gives better insight into the potential of traditional medicine and the critical role of microfluidics in the drug development process. In addition, the emergence of new ideas and applications will bring about further advances in the field of drug development. [ABSTRACT FROM AUTHOR]

In the present work, 27 triterpene derivatives have been subjected to 3D-QSAR, ADME-Tox, and molecular docking for their insecticidal activity. The selected derivatives are previously semi-synthesized based on compounds obtained from Euphorbia resinifera and Euphorbia officinarum latex. The in silico studies were used to predict and to evaluate the antibacterial and insecticidal properties of the 3D structure of triterpene derivatives. The 3D-QSAR models are developed using CoMFA and CoMSIA techniques, and they have showed excellent statistical results (R2 = 0.99; Q2 = 0.672; R2pred = 0.91 for CoMFA and R2 = 0.97; Q2 = 0.61; R2pred = 0.94 for CoMSIA). The results indicate that the built models are able to describe the relationship between the structure of triterpene derivatives and the pLD50 bioactivity. Based on contour maps obtained from CoMFA and CoMSIA models, 38 new molecules are designed and their pLD50 activities are predicted. The drug-like and ADME-Tox properties of the molecule designed are examined and led to the selection of four molecules (55, 56, 59, 64) as promising antibacterial and insecticidal agents. Compounds 55, 56, 59, and 64 are able to inhibit the MurE (PDB code: 1E8C) and EcR (PDB code: 1R20) proteins involved in the process of antibacterial and insecticidal activities. This hypothesis is confirmed by the implementation of a molecular docking test. This test predicted the most important referential interactions that occur between the structure of triterpene derivatives and the targeted receptors. Among the four docked molecules, three molecules (55, 56, and 59) showed greater stability than the reference molecule 16 inside the MurE and EcR receptors pocket. Therefore, the structure of the three new triterpene derivatives can be adopted as reference for the synthesis of antibacterial drugs and also in the development of insecticides. [ABSTRACT FROM AUTHOR]

N-dealkylation, the removal of an N-alkyl group from an amine, is an important chemical transformation which provides routes for the synthesis of a wide range of pharmaceuticals, agrochemicals, bulk and fine chemicals. N-dealkylation of amines is also an important in vivo metabolic pathway in the metabolism of xenobiotics. Identification and synthesis of drug metabolites such as N-dealkylated metabolites are necessary throughout all phases of drug development studies. In this review, different approaches for the N-dealkylation of amines including chemical, catalytic, electrochemical, photochemical and enzymatic methods will be discussed. [ABSTRACT FROM AUTHOR]

Background: Quinolines have demonstrated various biological activities such as antimalarial, antibacterial and anticancer. Hence, compounds with such scaffold have been used as lead in drug development. This project is, therefore, aimed to synthesis and evaluates some biological activities of quinoline analogs. Methods: 2-Chloro-7-fluoroquinoline-3-carbaldehydes were synthesized by the application of Vilsmeier–Haack reaction. The chlorine in the fluoroquinoline-3-carbaldehyde was replaced with various nucleophiles. The aldehyde functional group was also converted to carboxylic acid and imine groups using oxidizing agent and various amines, respectively. The structures of the compounds synthesized were characterized by spectroscopic methods. Disc diffusion and DPPH assays were used to evaluate the antibacterial and antioxidant activities, respectively. The in silico molecular docking analysis of the synthesized compounds were done using AutoDock Vina against E. coli DNA Gyrase B and human topoisomerase IIα. The drug likeness properties were assessed using SwissADME and PreADMET. Results: Nine novel quinoline derivatives were synthesized in good yields. The in vitro antibacterial activity of the synthesized compounds was beyond 9.3 mm inhibition zone (IZ). Compounds 4, 5, 6, 7, 8, 10, 15, and 16 exhibited activity against E. coli, P. aeruginosa, S. aureus and S. pyogenes with IZ ranging from 7.3 ± 0.67 to 15.3 ± 0.33 mm at 200 μg/mL. Compound 9 displayed IZ against three of the bacterial strains except S. aureus. The IC50 for the radical scavenging activity of the synthesized compounds were from 5.31 to 16.71 μg/mL. The binding affinities of the synthesized compounds were from − 6.1 to − 7.2 kcal/mol against E. coli DNA gyrase B and − 6.8 to − 7.4 kcal/mol against human topoisomerase IIα. All of the synthesized compounds obeyed Lipinski's rule of five without violation. Conclusion: Compounds 4, 5, 6, 7, 8, 10, 15, and 16 displayed activity against Gram positive and Gram negative bacterial strains indicating that these compounds might be used as broad spectrum bactericidal activity. Compound 8 (13.6 ± 0.22 mm) showed better IZ against P. aeruginosa compared with ciprofloxacin (10.0 ± 0.45 mm) demonstrating the potential of this compound as antibacterial agent against this strain. Compounds 5, 6, 7, 8, 9 and 10 showed comparable binding affinities in their in silico molecular docking analysis against E. coli DNA gyrase B. All of the synthesized compounds also obeyed Lipinski's rule of five without violation which suggests these compounds as antibacterial agents for further study. Compounds 7 and 8 were proved to be a very potent radical scavenger with IC50 values of 5.31 and 5.41 μg/mL, respectively. Compound 5, 6, 8, 10 and 16 had comparable binding affinity against human topoisomerase IIα suggesting these compounds as a possible candidate for anticancer drugs. [ABSTRACT FROM AUTHOR]

The term traditional medicine is used to explain the traditional medical practice that has been in existence even before the advent of modern medicine. It is widely accepted and used in the prevention and treatment of physical and mental treatment as well as a social imbalance. There seem to be limits to the development of new drugs based exclusively on modern technologies. Since the 1980s, the pharmaceutical industry has tended to use high throughput synthesis and drug development based on combinatorial chemistry in developing new drugs/medicines. However, significant efforts in this direction have not resulted in the drug's expected productivity. Some large pharmaceutical companies are facing significant challenges in developing new products. Our study was to evaluate the aged population's insight regarding traditional medicine and modern medicine. It was a prospective study with duration of three months from February 2021 to April 2021. The study was conducted in the population in and around Vijayawada. A total of 400 samples were collected. Generally, nowadays people are not aware of traditional medicines even if they are inexpensive when compared to modern drugs. So we conducted this study to know the knowledge of aged peoples on traditional medicine and to know their perception regarding both modern medicine and traditional medicine. [ABSTRACT FROM AUTHOR]

[Display omitted] • Commentary on when and how drug metabolite synthesis fits into the overall drug development pathway and the influence of regulatory guidelines. • The tool box of techniques used by a specialist contract research organisation to make phase I and II metabolites. • Case studies illustrating recent examples, including the significance of selected drug metabolites and their synthesis. The drug discovery and development process encompasses the interrogation of metabolites arising from the biotransformation of drugs. Here we look at why, when and how metabolites of small-molecule drugs are synthesised from the perspective of a specialist contract research organisation, with particular attention paid to projects for which regulatory oversight is relevant during this journey. To illustrate important aspects, we look at recent case studies, trends and learnings from our experience of making and identifying metabolites over the past ten years, along with with selected examples from the literature. [ABSTRACT FROM AUTHOR]

This proposal would synthesize the work on arylpiperazine derivatives because aryl piperazine has the largest amount of positive effects on the biological screen of the heterocyclic and its congeners as the most important building blocks for drug development. Two separate series of N-(4-(benzo[d]oxazol-2-yl)phenyl)-2-[4-(arylsubstituted) piperazines-1-yl]acetamide and 2-[4-(arylsubstitution)piperazine-1-yl]acetamide are synthesized and are distinguished by various substituents of aryl piperzine, such as a melting point determination and chromatography in thin-scale (TLC). Two different series are also performed, synthesized and characterized. Its spectroscopic characterisation was undertaken for their structure elucidation. Followed by this, synthesized drugs can be pharmacologically assessed. In pharmacological research, the inhibition of 5-hydoxytraphtophen (5-HTP) mediated head twitches, the inhibition of the behaviour, and the activation of cataleptic disorder are determined by the behaviours. [ABSTRACT FROM AUTHOR]

S -adenosylmethionine (SAM) is ubiquitous in living organisms and is of great significance in metabolism as a cofactor of various enzymes. Methyltransferases (MTases), a major group of SAM-dependent enzymes, catalyze methyl transfer from SAM to C, O, N, and S atoms in small-molecule secondary metabolites and macromolecules, including proteins and nucleic acids. MTases have long been a hot topic in biomedical research because of their crucial role in epigenetic regulation of macromolecules and biosynthesis of natural products with prolific pharmacological moieties. However, another group of SAM-dependent enzymes, sharing similar core domains with MTases, can catalyze nonmethylation reactions and have multiple functions. Herein, we mainly describe the nonmethylation reactions of SAM-dependent enzymes in biosynthesis. First, we compare the structural and mechanistic similarities and distinctions between SAM-dependent MTases and the non-methylating SAM-dependent enzymes. Second, we summarize the reactions catalyzed by these enzymes and explore the mechanisms. Finally, we discuss the structural conservation and catalytical diversity of class I-like non-methylating SAM-dependent enzymes and propose a possibility in enzymes evolution, suggesting future perspectives for enzyme-mediated chemistry and biotechnology, which will help the development of new methods for drug synthesis. Non-methylating S -adenosylmethionine (SAM)-dependent enzymes share similar core domain with methyltransferases but catalyze versatile reactions through different mechanisms involved in building significant bioactive compounds. The review on structure, catalytic mechanism and evolution of these enzymes may help the development of enzyme-mediated biotechnologies and new methods for drug synthesis. [Display omitted] • SAM-dependent enzymes have long been a hot topic in biomedical research. • SAM-dependent enzymes share structural similarity but can catalyze versatile reactions involved in building important bioactive compounds. • The review focuses on SAM-dependent enzymes that catalyze non-methylation. [ABSTRACT FROM AUTHOR]

The present communication reports the improved synthesis of an API molecule Quetiapine established by using R software for quality by design which improved the yield. An optimized synthesis by screening different solvents and bases was proposed. Finally better conditions for preparation of quetiapine were established using Quality by design. [ABSTRACT FROM AUTHOR]

The evaluation of drug metabolites is compulsory during drug development. Since recently, bacterial cytochromes P450 and their mutated variants have attracted considerable interest as an alternative to hepatic P450s for the synthesis of human drug metabolites. Thus, straightforward screening approaches are required that enable rapid identification and evaluation of drug‐metabolizing bacterial P450s with different product selectivities. Herein, we report a two‐step screening method for discovery and characterization of new P450s from actinomycetes that enable oxidation of various drugs. In the first step, substrate profiling with three structurally different model drugs, ritonavir, testosterone, amitriptyline, allowed us to select CYP105D and CYP107Z from Streptomyces platensis DSM 40041 that accepted all model substrates and produced human‐like drug metabolites. In the second step, activity tests with an array of 25 structurally‐related molecules and derivatives of the three model compounds revealed a correlation between structural variations in the target drugs and the enzyme chemo‐ and regioselectivity. [ABSTRACT FROM AUTHOR]

Organic catalysis: The activation of ubiquitous but unreactive carbon‐hydrogen bonds has emerged as an environmentally friendly alternative to traditional techniques of molecular synthesis. This strategy can be used to minimise toxic by‐products and develop new functional materials, which may also be beneficial in applications such as polymer synthesis and drug development, among others. [ABSTRACT FROM AUTHOR]

The anti-inflammatory and antinociceptive activity of S(+)-2-amino-2-(benzylamino) propanoic acid (Npaa 34), R(-)-2-amino-2-(benzylamino) propanoic acid (Npaa 35), S(-)-2- amino-2-methyl-3-phenyl propanoic acid (Npaa 36), S(-)-2-amino-3-(4'-fluoro) phenylpropanoic acid (Npaa 38) and their non-selective COX inhibiting properties demonstrated in our previous studies approved that synthesized α-amino-aryl propanoic acid derivatives could serve as a potential source for discover and development of new non-steroidal anti-inflammatory drugs. In view of this the main purpose of presented study was investigation of testified compounds toxicity as an important drugs development step -- safety testing. In vitro cytotoxicity by flow cytometry and in vivo toxicity study for determination on certain toxicometry parameters and hazard class assessment were carried out. Obtained results indicated that investigated compound Npaa 38 can be classified to the Category 3 (LD50 > 300 mg/kg) and Npaa 34 to the Category 4 (LD50 > 2000 mg/kg) by Globally harmonized system of classification and labeling of chemicals and third class of the moderately toxic chemicals according WHO classification (LD50 value of Npaa 38 for rat administrated per os was 1300±102 mg/kg and for Npaa 34 was more than 2000 mg/kg). In a single application of the compounds to the intact rats' skin irritating effect was not revealed while they have a moderate irritant effect on the mucous membranes of rabbit's eyes. according to flow cytometry data, the S(+)-2-amino-2- (benzylamino) propanoic acid (Npaa 34) and S(-)-2-amino-2-methyl-3-phenyl propanoic acid (NPAA 36) do not display cytotoxicity. It was demonstrated, that in 500 μM concentrations the total number of peripheral blood mononuclear cells (pBMC) and monocytes was not changed significantly in presence of investigated non protein amino acids. [ABSTRACT FROM AUTHOR]

Drug discovery and development are among the most important translational science activities that contribute to human health and wellbeing. However, the development of a new drug is a very complex, expensive, and long process which typically costs 2.6 billion USD and takes 12 years on average. How to decrease the costs and speed up new drug discovery has become a challenging and urgent question in industry. Artificial intelligence (AI) combined with new experimental technologies is expected to make the hunt for new pharmaceuticals quicker, cheaper, and more effective. We discuss here emerging applications of AI to improve the drug discovery process. AI has enormous potential to revolutionize drug discovery. Computational prediction of atomic and molecular properties is the foundation of most de novo design strategies. Machine learning, a branch of AI, can now predict the physical and chemical properties of small molecules at quantum mechanics-level accuracy with much lower time-cost. AI is also able to search for correlations between molecular representations and biological and toxicological activities. AI-based algorithms are also being developed to efficiently probe the pathways of synthesis of novel drug candidates. In combination with robotic platforms, the chemical space for novel reactions can be explored by learning from automated analysis of reaction feasibility. [ABSTRACT FROM AUTHOR]

Abstract A series of new 4-hydroxycoumarin platinum(IV) complexes were designed, synthesized and evaluated as antitumor agents. All the title compounds display moderate to effective antitumor activities toward the tested cell lines and two prominent compounds were screened out with activities comparable to cisplatin and oxaliplatin. The mechanism investigation demonstrates that the platinum(IV) compounds could be reduced to bivalence and exert significant genotoxicity to tumor cells. Meanwhile the coumarin moiety endows the title compounds with cyclooxygenase inhibitory competence which might favour the reduction of tumor-related inflammation and further influence tumor proliferation. The coumarin platinum(IV) complex could effectively induce apoptosis of SKOV-3 cells through up-regulating the expression of caspase3 and caspase9. Furthermore, the conversion of platinum(II) drugs to platinum(IV) form via the conjunction with 4-hydroxycoumarin enhances the drug uptake in whole cells and DNA simultaneously. Moreover, the 4-hydroxycoumarin platinum(IV) complex could combine with human serum albumin via van der Waals force and hydrogen bond, which would influence their transport and bioactivities in vivo. Graphical abstract New 4-hydroxycoumarin platinum(IV) hybrids were designed, synthesized and evaluated as antitumor agents. The platinum(IV) core could be reduced to bivalence and display significant genotoxicity. Meanwhile the coumarin group endows the title compounds with cyclooxygenase inhibitory competence which would favour of reducing tumor-related inflammation and further influencing tumor proliferation. Unlabelled Image Highlights • Development of 4-hydroxycoumarin platinum(IV) hybrids as antitumor agents • Antitumor activities were evaluated and the action mechanism was investigated. • Active apoptotic pathway by up-regulating the expression of caspase3 and caspase9 • Exert higher accumulation in both whole cells and DNA than platinum(II) drugs • Combine with human serum albumin effectively by noncovalent forces [ABSTRACT FROM AUTHOR]

Abstract Multi-drug resistance (MDR) to anticancer drugs is the primary impediment to successful treatment of cancer. Hunting for new compounds with potent anti-MDR activity is an effectual approach to conquer cancer drug resistance. In this work, 33 new sulfur-containing 1,4-naphthoquinone oxime derivatives were prepared and investigated for their cytotoxicity against a panel of tumor cell lines and fibroblast normal cell line. Cell-based assay showed that most of target compounds displayed more potent cytotoxic potency than positive controls. Meanwhile, all of compounds were non-toxic to normal cells. More importantly, the cytotoxic activity of these oxime derivatives toward drug-resistant cancer cell lines was found to be much stronger than that toward drug-susceptible cell lines (anti-drug resistance coefficient (ADRC) > 1). Of these, compound 12 m was identified as the most effective molecule with IC50 values in the range of 0.29 ± 0.01 to 1.33 ± 0.05 μM toward MDR sublines. Further mechanism studies demonstrated that 12 m could inhibit colony formation, cause G1 phase arrest and promote cell apoptosis mediated by augmenting Bax/Bcl-2 ratio of Bel7402/5-FU cells. Our findings provide promising start points for development of sulfur-containing 1,4-naphthoquinone oxime derivatives as potential anti-MDR agents. Graphical abstract Image 1 Highlights • Thirty-three 1,4-naphthoquinone oxime derivatives were synthesized and investigated for their cytotoxicity. • Most oxime derivatives exerted more potent activity than positive controls. • Anti-drug resistance coefficients (ADRC) of 1,4-naphthoquinone oxime derivatives > 1 were first discovered. • 12m was found to be the most active to MDR sublines with IC 50 values ranging from 0.29 ± 0.01 to 1.33 ± 0.05 μM. • Preliminary anti-MDR mechanisms of 12m were investigated. [ABSTRACT FROM AUTHOR]

Abstract A series of reduced lipophilic N-benzylic imidazo[1,2-a]pyridine carboxamides (IPAs) with various side chains were designed and synthesized as new anti-TB agents in this work. Five derivatives A2 , A3 , A4 , B1 and B9 exhibit excellent in vitro activity (MIC: < 0.035 μM) against the drug susceptive Mycobacterium tuberculosis H37Rv strain and two clinically isolated multidrug-resistant strains, and acceptable PK properties. Compound B1 bearing a cyclohexylmethyl piperazine moiety, the same side chain of PBTZ169, was found to have obviously greater AUC 0-∞ and C max than Q203 and PBTZ169, suggesting its promising potential to be a lead compound for future antitubercular drug discovery. Graphical abstract Image 1 Highlights • Novel reduced lipophilic imidazo[1,2-a]pyridine-3-carboxamide derivatives were synthesized. • SARs were summarized for further antitubercular agents discovery. • Anti-MDR-TB activity, cytotoxicity and PK properties were investigated. • Compound B1 may serve as a new and promising candidate for further study. [ABSTRACT FROM AUTHOR]

Abstract Despite new agent development and short-term benefits in patients with colorectal cancer (CRC), metastatic CRC cure rates have not improved due to high rates of 5-fluorouracil (5-FU)/leucovorin/oxaliplatin (FOLFOX)-resistance and a clinical therapeutic plateau. At the same time, this treatment regime leads to significant toxicity, cost, and patient inconvenience. Drug-resistance is linked to CRC stem cells, which are associated with the epidermal-to-mesenchymal transition (EMT) pathway. Thus, to optimally treat CRC, a therapy that can target the cell survival and EMT pathways in both CRC bulk and stem cell populations is critical. We recently identified a novel small molecule NSC30049 (7a) that is effective alone, and in combination potentiates 5-FU-mediated growth inhibition of CRC bulk, FOLFOX-resistant, and CRC stem cells both in vitro and in vivo models. In the present study, we report the synthesis and anti-CRC evaluation of several stable and effective 7a analogs. ASR352 (7b) was identified as one of the equipotent 7a analogs that inhibited the growth of CRC bulk cells, sensitized FOLFOX-resistant cells, and reduced the sphere formation capacity of CRC stem cells. It appears that the complex mechanism of cytotoxicity for 7b includes abrogation of 5-FU-induced the S phase, reduction of the phosphorylation of Chk1 at S317P, S345P and S296P, increased γH2AX staining, activation of caspase 3/PARP1 cleavage, and enhancement of Bax/Bcl2 ratio. Further 7b -mediated reduced phosphorylation of Chk1 was an indirect effect, since it did not inhibit Chk1 activity in an in vitro kinase assay. Our findings suggest that 7b as a single agent, or in combination with 5-FU can be developed as a therapeutic agent in CRC bulk, FOLFOX-resistant, and CRC stem cell populations for unmanageable metastatic CRC conditions. Graphical abstract Image 1 Highlights • Novel analogs of orphan molecule NSC30049 were synthesized. • ASR352 (7b) was identified as most cytotoxic to colorectal cancer (CRC) cells. • 7b inhibited the growth of CRC bulk and sensitized FOLFOX-resistant cells. • 7b reduced sphere formation capacity of CRC stem cells. • 7b abrogated 5-FU-induced the S phase and inhibited the phosphorylation of Chk1. [ABSTRACT FROM AUTHOR]

Emergence of multidrug-resistant bacteria forces us to explore new therapeutic strategies, and proteins involved in key metabolic pathways are promising anti-bacterial targets. Bifunctional flavin-adenine dinucleotide (FAD) synthetases (FADS) are prokaryotic enzymes that synthesise the flavin mononucleotide (FMN) and FAD cofactors. The FADS from the human pathogen Streptococcus pneumoniae (SpnFADS)-causative agent of pneumonia in humans − shows relevant catalytic dissimilarities compared to other FADSs. Here, by integrating thermodynamic and kinetic data, we present a global description of the riboflavin kinase activity of SpnFADS, as well as of the inhibition mechanisms regulating this activity. Our data shed light on biophysical determinants that modulate species-specific conformational changes leading to catalytically competent conformations, as well as binding rates and affinities of substrates versus products. This knowledge paves the way for the development of tools − that taking advantage of the regulatory dissimilarities during FMN biosynthesis in different species − might be used in the discovery of specific anti-pneumococcal drugs. [ABSTRACT FROM AUTHOR]

Triple-negative breast cancer (TNBC) is a highly aggressive malignancy with limited treatment options due to its heterogeneity and the lack of well-defined molecular targets. In our endeavour towards the development of novel anti-TNBC agents, herein we report a one-pot three-component synthesis of novel spirooxindoles 6a-p, and evaluation of their potential anti-proliferative activity towards TNBC MDA-MB-231 cells. Spirooxindoles 6a, 6e and 6i emerged as the most potent analogues with IC50 = 6.70, 6.40 and 6.70 µM, respectively. Compounds 6a and 6e induced apoptosis in MDA-MB-231 cells, as evidenced by the up-regulation of the Bax and down-regulation of the Bcl-2, besides boosting caspase-3 levels. Additionally, 6e displayed significant increase in the percent of annexin V-FITC positive apoptotic cells from 1.34 to 44%. Furthermore, spirooxindoles 6e and 6i displayed good inhibitory activity against EGFR (IC50 = 120 and 150 nM, respectively). Collectively, these data demonstrated that 6e might be a potential lead compound for the development of effective anti-TNBC agents. [ABSTRACT FROM AUTHOR]

Drug discovery and development is a long-lasting process in which many challenges have to be addressed at every stage, from the discovery of the target biomolecule to the commercialization of the discovered drugs and its quality control. From thousands of hits identified during early drug discovery only one drug is eventually efficient and safe enough to be commercialized. This high rejection rate, especially during preclinical and clinical studies have led to an exponential increase of costs to develop new medicines thereby strongly impacting healthcare systems. In this context, miniaturized devices have the potency to significantly reduce the cost and the time needed to develop new therapeutics by streamlining drug development and rejecting drug candidates earlier in the process prior to costly animal and human trials. In this review, we present recent advances involving miniaturized technologies in the field of drug discovery such as target discovery, drug screening, drug synthesis and formulation, in-vitro and in-vivo testing and quality control. We discuss opportunities enabled by miniaturized devices but also their limitations and challenges that need to be resolved in order to spread their use in the pharmaceutical industries. [ABSTRACT FROM AUTHOR]

Fourteen novel amino-quinoline-5,8-dione derivatives ( 6a-h and 7a-h ) were designed and synthesized by coupling different alkyl- or aryl-amino fragments at the C6- or C7-position of quinoline-5,8-dione. All target compounds showed antiproliferative potency in the low micromolar range in both drug sensitive HeLaS3 and multidrug resistant KB-vin cell lines. Compounds 6h , 6d , 7a , and 7d exhibited more potent antiproliferative effects than the other compounds. Especially, compounds 6d and 7d displayed NQO1-dependent cytotoxicity and competitive NQO1 inhibitory effects in both drug sensitive HeLaS3 and multidrug resistant KB-vin cell lines. Furthermore, compounds 6h , 6d , 7a , and 7d induced a dose-dependent lethal mitochondrial dysfunction in both drug sensitive HeLaS3 and multidrug resistant KB-vin cells by increasing intracellular reactive oxygen species (ROS) levels. Notably, compound 7d selectively inhibited cancer cells, but not non-tumor liver cell proliferation in vitro , and significantly triggered HeLaS3 cell apoptosis by regulating apoptotic proteins of Bcl-2, Bax, and cleaved caspase-3 in a dose-dependent manner. Our findings suggest that these novel C6- or C7-substituted amino-quinoline-5,8-dione derivatives, such as 7d , could be further developed in the future as potent and selective antitumor agents to potentially circumvent multi-drug resistance (MDR). [ABSTRACT FROM AUTHOR]

N -oxide derivatives 5(a – b) , 8(a – b) , and 11(a – c) were designed, synthesized and evaluated in vitro and in vivo as potential drugs that are able to ameliorate sickle cell disease (SCD) symptoms. All of the compounds demonstrated the capacity to releasing nitric oxide at different levels ranging from 0.8 to 30.1%, in vivo analgesic activity and ability to reduce TNF-α levels in the supernatants of monocyte cultures. The most active compound ( 8b ) protected 50.1% against acetic acid-induced abdominal constrictions, while dipyrone, which was used as a control only protected 35%. Compounds 8a and 8b inhibited ADP-induced platelet aggregation by 84% and 76.1%, respectively. Both compounds increased γ-globin in K562 cells at 100 μM. The mechanisms involved in the γ-globin increase are related to the acetylation of histones H3 and H4 that is induced by these compounds. In vitro , the most promising compound ( 8b ) was not cytotoxic, mutagenic and genotoxic. [ABSTRACT FROM AUTHOR]

Over the past few decades, various computational methods have become increasingly important for discovering and developing novel drugs. Computational prediction of chemical reactions is a key part of an efficient drug discovery process. In this review, we discuss important parts of this field, with a focus on utilizing reaction data to build predictive models, the existing programs for synthesis prediction, and usage of quantum mechanics and molecular mechanics (QM/MM) to explore chemical reactions. We also outline potential future developments with an emphasis on pre-competitive collaboration opportunities. [ABSTRACT FROM AUTHOR]

The synthesis of stable isotope labeled (SIL) complex drug molecules with a ≥3 mass unit increase from the parent compound is essential for drug discovery and development. Typical approaches that rely on 2H, 13C, and 15N isotopes can be very challenging or even intractable, and can delay the drug development process. This work introduces a new concept for the synthesis of labeled compounds that relies on the use of 34S. The synthetic utility of 34S was demonstrated with the efficient synthesis of [34S]phosphorothioates [34S2]‐PS‐ODNs‐TTT and [13C, 15N, 34S]‐ceftolozane. In addition, a procedure for the direct oxidation of phosphites to [34S]phosphorothioates using elemental 34S without isotope dilution was developed. [ABSTRACT FROM AUTHOR]

Diversity-oriented synthesis of derivatives of natural products is an important approach for the discovery of novel drugs. In this paper, a series of novel 3,4-diaryl-1 H -pyrazoles and 3,5-diaryl-1 H -pyrazoles derivatives were synthesized through the one-pot reaction of flavones and isoflavones with the hydrazine hydrate and substituted hydrazine hydrate. Some of these novel compounds exhibited antifungal effects against Candida albicans SC5314, and displayed more potent inhibitory activities against the efflux-pump-deficient strain DSY654. In addition, compounds 25 , 28 and 32a displayed outstanding reversal activity of azole resistance against clinical azole-resistant Candida albicans in combination with fluconazole (FLC), with FICI values ranging from 0.012 to 0.141. The preliminary structure-activity relationship (SAR) of these compounds was also discussed. In conclusion, this study provides several novel agents that displayed potent antifungal activities alone or together with fluconazole, which makes progress for development of antifungal drugs. [ABSTRACT FROM AUTHOR]

Today’s highly competitive pharmaceutical industry is in dire need of an accelerated transition from the drug development phase to the drug production phase. At the heart of this transition are chemical reactors that facilitate the synthesis of active pharmaceutical ingredients (APIs) and whose design can affect subsequent processing steps. Inspired by this challenge, we present a model-based approach for systematic reactor design. The proposed concept is based on the elementary process functions (EPF) methodology to select an optimal reactor configuration from existing state-of-the-art reactor types or can possibly lead to the design of novel reactors. As a conceptual study, this work summarizes the essential steps in adapting the EPF approach to optimal reactor design problems in the field of API syntheses. Practically, the nucleophilic aromatic substitution of 2,4-difluoronitrobenzene was analyzed as a case study of pharmaceutical relevance. Here, a small-scale tubular coil reactor with controlled heating was identified as the optimal set-up reducing the residence time by 33% in comparison to literature values. [ABSTRACT FROM AUTHOR]

Finding and optimising of synthesis processes for active pharmaceutical ingredients (API) is time consuming. In the finding phase, established methods for synthesis, purification and formulation are used to achieve a high purity API for biological studies. For promising API candidates, this is followed by pre-clinical and clinical studies requiring sufficient quantities of the active component. Ideally, these should be produced with a process representative for a later production process and suitable for scaling to production capacity. This work presents an overview of different approaches for process synthesis based on an existing lab protocol. This is demonstrated for the production of the model drug 4,5,6,7-tetrabromo-2-(1 H -imidazol-2-yl) isoindolin-1,3-dione (TBID). Early batch synthesis and purification procedures typically suffer from low and fluctuating yields and purities due to poor process control. In a first step the literature synthesis and purification procedure was modified and optimized using solubility measurements, targeting easier and safer processing for consecutive studies. [ABSTRACT FROM AUTHOR]

HxTfA 4 is a fluorescent analog of a potent cytotoxic and antimalarial agent, TfA 3 , which is currently being investigated for the development of an antimalarial vaccine, PlasProtect®. HxTfA contains a p-anisylbenzimidazole or Hx moiety, which is endowed with a blue emission upon excitation at 318 nm; thus enabling it to be used as a surrogate for probing the cellular fate of TfA using confocal microscopy, and addressing the question of nuclear localization. HxTfA exhibits similar selectivity to TfA for A-tract sequences of DNA, alkylating adenine-N3, albeit at 10-fold higher concentrations. It also possesses in vitro cytotoxicity against A549 human lung carcinoma cells and Plasmodium falciparum. Confocal microscopy studies showed for the first time that HxTfA, and by inference TfA, entered A549 cells and localized in the nucleus to exert its biological activity. At biologically relevant concentrations, HxTfA elicits DNA damage response as evidenced by a marked increase in the levels of γH2AX observed by confocal microscopy and immunoblotting studies, and ultimately induces apoptosis. [ABSTRACT FROM AUTHOR]

Need for new drugs to fight against tuberculosis (TB) is increasing day by day. In the present work we have taken a spiro compound (GSK 2200150A) reported by GSK as a lead and we modified the structure of the lead to study the antitubercular activity. For structure activity profiling twenty-one molecules have been synthesized, characterized and evaluated for their antimycobacterial potency against both active and dormant TB. Compound 06 , 1-((4-methoxyphenyl)sulfonyl)-4′,5′-dihydrospiro[piperidine-4,7′-thieno[2,3- c ]pyran] was found to be the most potent compound (MIC: 8.23 µM) in active TB and was less effective than the lead but more potent than standard first line drug ethambutol. It was also found to be more efficacious than Isoniazid and Rifampicin and equipotent as Moxifloxacin against dormant Mycobacterium tuberculosis (MTB). Compound 06 also showed good inhibitory potential against over expressed latent MTB enzyme lysine ε-amino transferase with an IC 50 of 1.04 ± 0.32 µM. This compound is a good candidate for drug development owing to potential against both active and dormant stages of MTB. [ABSTRACT FROM AUTHOR]

A synthetic method for 7,10-dimethyl-2-thioxo-2,3-dihydro-1H-spiro[benzo[h]quinazoline-5,1′-cyclopentane]- 4(6H)-one from ethyl 4′-amino-5′,8′-dimethyl-1′-H-spiro[cyclopentane-1,2′-naphthalene]-3′-carboxylate was developed. Reaction of the spirobenzo[h]quinazoline with various alkyl(benzyl) halides synthesized a novel series of spirobenzo[h]quinazolines containing methyl substituents on the benzene ring. The antidepressant, anticonvulsant, and antibacterial activities of the synthesized spirobenzo[h]quinazolines were investigated. [ABSTRACT FROM AUTHOR]

Neuropeptide Y2 receptor (Y2R) agonism is an important anorectic signal and a target of antiobesity drug discovery. Recently, we synthesized a short-length Y2R agonist, PYY-1119 (4-imidazolecarbonyl-[ d -Hyp 24 ,Iva 25 ,Pya(4) 26 ,Cha 27,36 ,γMeLeu 28 ,Lys 30 ,Aib 31 ]PYY(23–36), 1 ) as an antiobesity drug candidate. Compound 1 induced marked body weight loss in diet-induced obese (DIO) mice; however, 1 also induced severe vomiting in dogs at a lower dose than the minimum effective dose administered to DIO mice. The rapid absorption of 1 after subcutaneous administration caused the severe vomiting. Polyethylene glycol (PEG)- and alkyl-modified derivatives of 1 were synthesized to develop Y2R agonists with improved pharmacokinetic profiles, i.e., lower maximum plasma concentration (C max ) and longer time at maximum concentration (T max ). Compounds 5 and 10 , modified with 20 kDa PEG at the N-terminus and eicosanedioic acid at the Lys 30 side chain of 1 , respectively, showed high Y2R binding affinity and induced significant body weight reduction upon once-daily administration to DIO mice. Compounds 5 and 10 , with their relatively low C max and long T max , partially attenuated emesis in dogs compared with 1 . These results indicate that optimization of pharmacokinetic properties of Y2R agonists is an effective strategy to alleviate emesis induced by Y2R agonism. [ABSTRACT FROM AUTHOR]

The scarcity of complex intermediates in pharmaceutical research motivates the pursuit of reaction optimization protocols on submilligram scales. We report here the development of an automated flow-based synthesis platform, designed from commercially available components, that integrates both rapid nanomole-scale reaction screening and micromole-scale synthesis into a single modular unit. This system was validated by exploring a diverse range of reaction variables in a Suzuki-Miyaura coupling on nanomole scale at elevated temperatures, generating liquid chromatography-mass spectrometry data points for 5760 reactions at a rate of >1500 reactions per 24 hours. Through multiple injections of the same segment, the system directly produced micromole quantities of desired material. The optimal conditions were also replicated in traditional flow and batch mode at 50- to 200-milligram scale to provide good to excellent yields. [ABSTRACT FROM AUTHOR]

Chemical manufacturing is often done at large facilities that require a sizable capital investment and then produce key compounds for a finite period. We present an approach to the manufacturing of fine chemicals and pharmaceuticals in a self-contained plastic reactionware device. The device was designed and constructed by using a chemical to computer-automated design (ChemCAD) approach that enables the translation of traditional bench-scale synthesis into a platform-independent digital code. This in turn guides production of a three-dimensional printed device that encloses the entire synthetic route internally via simple operations. We demonstrate the approach for the g-aminobutyric acid receptor agonist, (±)-baclofen, establishing a concept that paves the way for the local manufacture of drugs outside of specialist facilities. [ABSTRACT FROM AUTHOR]

Lysine specific demethylase 1 (LSD1), the first identified histone demethylase, plays an important role in epigenetic regulation of gene activation and repression, has been reported to be up-regulated and involved in numbers of solid malignant tumors. In this study, we identified a series of phenylalanyl hydrazones based LSD1 inhibitors, and the most potent one, compound 4q , can inactivate LSD1 with IC 50  = 91.83 nM. In cellular level, compound 4q can induce the accumulation of CD86 as well as H3K4me2, and inhibit gastric cancer cell proliferation by inactivating LSD1. Our findings indicated that compound 4q may serve as a potential leading compound to target LSD1 overexpressed gastric cancer. [ABSTRACT FROM AUTHOR]

The genotoxic impurities (GIs) are carcinogenic hence its management during synthesis of pharmaceuticals is very important to be detected even in trace level for the safe use of the drugs. The presence of drug substance/drug product DNA-reactive impurities poses a significant problem for drug regulators as well as industry. There are several regulatory guidelines and position papers focused on controlling the amount of impurities within the specified limits. The present compilation gives an account of updated information about GIs and reviews the regulatory aspects for GIs in active pharmaceutical ingredients/drug formulations. A detailed discussion about control strategies in the context of GIs is also described precisely. The analysis of GIs is a challenging and complex aspect of the drug development process. Control and determination of these impurities at ppm or ppb levels are significant challenges for analysts, therefore the approaches for the analysis of GIs have also been discussed. [ABSTRACT FROM AUTHOR]