Drugs and Drug Candidates

Background/Objectives: Pancreatic cancer is the seventh most lethal type of cancer in the world, and its treatment, which is largely inefficient, is based on surgery and/or non-specific chemotherapy. Its malignant features are characterized by complex cell signaling pathways, which can be used as targets for new drugs. Methods: In this study, glucal-based compounds were synthetized, with substitution based on fluorine, nitrogen and aromatic ring addition. The compounds were tested in the pancreatic cell culture Mia-PaCa-2 and cell viability was assessed, with further IC50 calculation, stability and selectivity. Molecular docking was performed to evaluate the probable molecular target for 5b and in silico physicochemical properties were determined. Results: One molecule, named 5b, with two fluorine atoms inserted in the aromatic ring, exerted potent inhibitory activity on cell growth (IC50 = 1.39 µM), which was selective for pancreatic cells. Through molecular docking studies, the compound was found to be positioned in the active site of JAK3, indicating inhibition of such protein, which has a role in tumoral cell growth. Moreover, 5b was stable for 24 months and had physicochemical properties to permeate cell membranes, good oral absorption, and low potential to cause toxicity. Conclusions: These data suggest that 5b can be druggable and can be considered as a prototype for a new course of treatment in pancreatic cancer. Full article

Background/Objectives: The thiazolo [5,4-d]pyrimidine scaffold is a class of drugs known for its anticancer, antitumor, anti-inflammatory, and antimicrobial properties. In this study, the electrochemical properties of novel thiazolo [5,4-d]pyrimidine derivatives and their interactions with DNA were characterized for the first time using voltammetric methods. Determining the interactions of new drug candidate molecules with DNA is crucial for drug development studies and is the main objective of this research. Methods: Both molecules were immobilized on the surface of the electrodes by passive adsorption, and their electrochemical properties were determined by voltammetric methods through reduction currents. Their interactions with DNA were carried out in the solution phase and examined by the changes in the oxidation peak potential and current of the guanine base. Results: For both molecules, it was determined that the electrochemical reduction processes are diffusion-controlled and irreversible, with an equal number of protons and electrons being transferred during this process. The detection limits for TP-NB (4-chloro-N-(5-chlorothiazolo [5,4-d]pyrimidin-2-yl)-3-nitrobenzamide) and TP-PC (1-(2-(4-(4-carbamoylpiperidin-1-yl)-3-nitrobenzamido)thiazolo [5,4-d]pyrimidin-5-yl)piperidine-4-carboxamide) were determined to be 12 µg/mL and 16 µg/mL, respectively. As a result of the interaction between both molecules with DNA, the guanine oxidation current decreased. It was found that TP-NB could act as an intercalator, while TP-PC could affect DNA electrostatically, both showing toxic effects on DNA. Conclusions: An electrochemical method was developed for the rapid, cost-effective, and sensitive detection of both molecules and their DNA interactions. Both compounds exhibited notable affinity towards DNA, as evidenced by significant changes in oxidation peak currents, shifts in peak potentials, and calculated toxicity values. These findings suggest their potential use as DNA-interacting drugs, such as anticancer and antimicrobial agents. Our study offers a quick, cost-effective, and reliable electrochemical approach for the evaluation of drug–DNA interactions. Full article

Engineered exosomes have emerged as transformative drug carriers, uniquely equipped to overcome biological barriers in central nervous system (CNS) disorders and cancer therapy. These natural extracellular vesicles, derived from cell membranes, offer inherent biocompatibility, low immunogenicity, and the ability to traverse physiological obstacles such as the blood–brain barrier (BBB) and dense tumor stroma. Recent advances in exosome engineering—including surface modification (e.g., ligand conjugation for receptor-mediated targeting) and cargo loading (siRNA, CRISPR-Cas systems, and chemotherapeutics)—have enhanced their precision and therapeutic utility. For CNS delivery, exosomes functionalized with brain-homing peptides (e.g., RVG or TfR ligands) have enabled the efficient transport of neuroprotective agents or gene-editing tools to treat Alzheimer’s disease or glioblastoma. In oncology, engineered exosomes loaded with tumor-suppressive miRNAs or immune checkpoint inhibitors exploit tumor microenvironment (TME) features, such as acidity or enzyme overexpression, for spatially controlled drug release. Furthermore, hybrid exosome–liposome systems and exosome–biomaterial composites are being explored to improve payload capacity and stability. Despite progress, challenges persist in scalable production, batch consistency, and regulatory standardization. This review critically evaluates engineering strategies, preclinical success, and translational hurdles while proposing innovations such as AI-driven exosome design and patient-derived exosome platforms for personalized therapy. By bridging nanotechnology and biomedicine, engineered exosomes can represent a paradigm shift in targeted drug delivery, offering safer and more effective solutions for historically intractable diseases. Full article

Background/Objectives: Bacterial infections remain among the top ten major public health concerns, contributing to a high number of incidences of disease and mortality worldwide, exacerbated by the rise of multidrug-resistant bacteria (MDRB). Consequently, it is crucial to develop novel antimicrobial strategies, including the use of functional nanoparticles. Gold nanoparticles (GNPs) have emerged as promising candidates due to their unique optical properties, particularly their ability to efficiently convert absorbed light into heat through the photothermal (PT) effect, which can be harnessed for bacteria eradication. Methods: Chitosan was modified with 3-mercaptopropionic acid to introduce sulfur groups, facilitating gold deposition onto chitosan nanoparticle (TCNPs) surface. The gold shell was subsequently formed via a seed-mediated method, wherein gold seeds were adsorbed onto TCNPs and further grown to form the shell. Photothermal effect on the bacterial viability was evaluated. Results: TCNPs with a size of 178 nm and spherical morphology were obtained. After the gold shell (TCNP@Au) exhibited a photothermal conversion efficiency of 31%, making them a promising photothermal agent for bacterial clearance. Notably, the viability of Escherichia coli was significantly reduced in the presence of TCNP@Au and was almost eradicated upon PT treatment. In contrast, TCNP@Aus were non-toxic for Staphylococcus aureus. Conclusions: TCNP@Au demonstrated favorable photothermal properties, presenting a novel nanoplatform for antibacterial applications, particularly against Gram-negative bacteria. However, further investigation is required to optimize the PT-based strategies against Gram-positive bacteria, such as S. aureus. Full article

Background: Metritis, a common postpartum uterine infection in bovines, poses substantial challenges in livestock management, including compromised fertility and economic losses. Poor uterine drug penetration and systemic side effects, necessitating innovative localised delivery systems and limiting current systemic antibiotic treatments. Aim: This study aimed to develop and evaluate the potential effect of the ofloxacin-loaded hydrogel as a localised drug delivery system to treat metritis in bovine. The focus was on achieving sustained drug release, enhanced antibacterial efficacy and reduced inflammation in the endometrium. Materials and Methods: The CS/PVA hydrogel was synthesised using a freeze–thaw method and further optimised for drug encapsulation efficiency (96.7 ± 2.1%), stability and biocompatibility. Physicochemical characterisation included swelling behaviour, mechanical properties and rheological analysis. In vitro drug release profiles in the simulated uterine fluid were assessed over 72 h and antibacterial activity was tested against common uterine pathogens such as Escherichia coli and S. aureus. In vivo studies were conducted on bovines diagnosed with endometritis to evaluate clinical recovery. Results: The SEM image of the ofloxacin-loaded CS/PVA hydrogel resulted in a smooth and porous structure demonstrating larger pore size than the blank. The rheological study suggested higher stability and elastic behaviour. Antibacterial assays on E. coli and S. aureus revealed significant inhibition zones, respectively, indicating potent efficacy. In vivo, evaluated on treated bovine, reduced bacterial loads were exhibited (2.86 × 10^5A CFU/mL → 6.37 × 10^2B CFU/mL), clinical improvement was marked and uterine inflammation was resolved. Conclusions: Ofloxacin-loaded hydrogels represent a promising localised treatment for bovine metritis, offering sustained antibacterial action and improved clinical outcomes. This approach addresses the limitations of systemic antibiotic therapies and provides a practical solution for enhanced veterinary care. Further studies are recommended to validate these findings in more extensive field trials and explore commercialisation potential. Full article

Background: The cholinergic hypothesis is an elementary approach employed for the research and drug discovery of novel anti-Alzheimer therapeutics. Method: In this context, the study focuses on synthesizing and evaluating a new series of chalcone derivatives (3a–3j) as multifunctional therapeutic agents, specifically investigating their antioxidant potential using the DPPH method with ascorbic acid as a standard. Ellman’s protocol for acetylcholinesterase inhibition assay was performed using donepezil as a standard, and computational insights were explored through molecular docking and ADME profiling. Results: Compounds 3a, 3d, 3e, 3f, and 3h exhibited excellent antioxidant activity compared to the standard. Most of the compounds exhibited moderate to good (3b, 3c, and 3h) AChE inhibitory activity. Molecular docking studies revealed conventional hydrogen bonding and π-π interactions with the enzyme’s active residues, facilitated by their electronegative groups and phenyl rings, respectively. In addition, a pharmacokinetic study was conducted using computational approach to assess druggability. The results demonstrated that compound 3b is an outstanding lead candidate with appreciable AChE inhibitory activity. Conclusions: The combined experimental and computational results of this study highlight the multifunctional nature of chalcone derivatives, suggesting their potential as promising therapeutic agents for the discovery of novel AChE inhibitors that could be employed in the management of Alzheimer’s disease and oxidative stress-related diseases. Full article

To date, numerous studies have emerged that indicate the possible role of epigenetic modulation in the development and progression of several diseases, including cancer. Epigenetic alterations participate in various steps of carcinogenesis. They play important regulatory roles in processes like cell division, proliferation, angiogenesis, and metastasis. Thus, epigenetic modifications such as DNA methylation, histone modifications, and non-coding RNAs serve as attractive and promising targets for cancer prevention and anti-cancer therapy. Epigenetic drugs or epi-drugs possess the ability to reverse many such epigenetic alterations and thus can help manage the clinical manifestations of cancer. Epigenetic drugs broadly target epigenetic modifications, including DNA methylation and histone post-translational modifications, to manifest their effects. Several naturally occurring as well as chemically synthesized compounds have been recognized as epigenetic drugs. Some of them are clinically approved, while many are in their preclinical and clinical trials. In this review, we aim to present a broad overview of the epigenetic modifications implicated in carcinogenesis. The review also compiles various epigenetic drugs that are approved for clinical practice, as well as those that are in the preclinical and clinical stages of investigation for anti-cancer therapy. Full article

Background/Objectives: Uveal melanoma (UM) is a rare but deadly ocular cancer. This review summarizes the characteristics of uveal melanoma and current therapeutic options for primary uveal melanoma and metastatic uveal melanoma, and introduces recent development of therapeutic strategies in preclinical animal studies and clinical trials. Methods: The literature search was conducted to identify relevant articles for UM studies. It was performed using PubMed for articles in English until March 2025. Information on clinical trials was also obtained from ClinicalTrial.gov. Results: Uveal melanoma originates from melanocytes, similar to skin melanoma. However, uveal melanoma has different mutations from skin melanoma. Thus, chemotherapy and immunotherapy, which are effective for skin melanoma, are ineffective for uveal melanoma. Current therapies for UM include radiation therapy, surgical resection, liver-directed therapies, and recently FDA-approved tebentafusp. Although a wide variety of available and newly developed therapies have been tested in clinical trials for UM, tebentafusp is the only FDA-approved therapy for metastatic UM. Given the excessive expression of vascular endothelial growth factor (VEGF) in UM patients with metastatic diseases, anti-VEGF strategies are being tested in clinical trials and pre-clinical animal models. Conclusions: This review summarizes an overview of current therapies and the development of therapeutic strategies in clinical trials and pre-clinical animal models. Despite successful control of primary tumors, 50% of UM patients still experience metastasis in the liver. Although tebentafusp improves the overall survival (OS) of a certain population of UM patients, robust strategies for preventing UM metastasis represent a critical unmet need. Further investigations of the heterogeneity of UM cells and mechanisms of UM metastasis are needed in the future. Full article

The COVID-19 pandemic, driven by the high transmissibility and immune evasion caused by SARS-CoV-2 and its variants (e.g., Alpha, Delta, Omicron), has led to massive casualties worldwide. As of November 2024, the International Committee on Taxonomy of Viruses (ICTV) has identified 14,690 viral species across 3522 genera. The increasing infectious and resistance to FDA and EMA-approved antivirals, such as 300-fold efficacy reduction in Nirmatrelvir against the SARS-CoV-2 3CLpro, highlight the need for mutation-stable antivirals, likewise targeting the essential host proteins like kinases, heat shock proteins, lipid metabolism proteins, immunological pathway proteins, etc. Unlike direct-acting antivirals, HDAs reduce the risk of resistance by targeting conserved host proteins essential for viral replication. The proposal for repurposing current FDA-approved drugs for host-directed antiviral (HDA) approach is not new, such as the Ouabain, a sodium-potassium ATPase inhibitor for herpes simplex virus (HSV) and Verapamil, a calcium channel blocker for influenza A virus (IAV), to name a few. Given the colossal potential of the mutation-stable HDA approach to exterminate the virus infection, it has been increasingly studied on SARS-CoV-2. This review aims to unravel the interaction between viruses and human hosts and their successfully proposed host-directed antiviral approach to provide insight into an alternative treatment to the rampant mutation in SARS-CoV-2. The benefits, limitations, and potential of host protein-targeted antiviral therapies and their prospects are also covered in this review. Full article

The mini-review highlights the innovative development of hetero-dinuclear metallic complexes, with a specific focus on zinc(II) metal centers arranged in a square-pyramidal configuration. The work presented, stemming from our research group in collaboration with others between the years 2020 and 2024, makes significant contributions to this area, emphasizing their potential applications in bioinorganic chemistry, particularly in the context of drug discovery. These advances not only expand the fundamental understanding of such complexes but also lay the groundwork for the design of novel hetero-dinuclear metallic compounds with therapeutic potential. The interaction of these complexes with biological systems and their implications for drug development are critical for future research in bioinorganic chemistry, offering new pathways for targeted treatments and molecular therapies. Full article

Background/Objectives: Sleep is essential to human health, playing a vital role in physical and mental well-being. Sleep disorders can lead to significant health complications, such as cardiovascular problems, diabetes, obesity, and depression. In Brazil, plants such as passionflower (Passiflora spp.), chamomile (Matricaria chamomilla L.) and mulungu (Erythrina spp.) are widely used in folk medicine for their sleep-promoting properties. This article reviews the existing literature on the sleep-promoting effects of these plants, focusing on the Brazilian context and popular knowledge of their use. Methods: An integrative literature review was conducted, including scientific articles in English and Portuguese from PubMed, Scielo and Google Scholar databases. Ethnobotanical studies documenting the traditional use of these plants in Brazil and clinical and preclinical research on their sleep-promoting effects were included. Results: The juice and infusion of the leaves and fruits of passionflower are mainly used to treat anxiety and insomnia, chamomile flower tea is used for its sedative effects, and mulungu bark decoctions are used for their sedative and anxiolytic properties. These popular uses are supported by scientific studies demonstrating the efficacy of these plants in treating insomnia, anxiety, and stress. Conclusions: The recognition of traditional knowledge and the inclusion of these plants in RENISUS highlights their importance for public health in Brazil. However, more rigorous clinical trials are needed to confirm their efficacy and safety and ensure their safe integration into modern medicine. Full article

Tanacetum balsamita is a perennial medicinal plant belonging to the Asteraceae family. The species bears a long history as a valuable traditional drug in different cultures, while it is an essential component in the traditional cuisine of several countries. In this context, our literature review aims at providing a comprehensive overview of T. balsamita, covering its traditional uses, phytochemistry, biological activities, and toxicity from 1983 to 2024. Methods: Various databases were used to collect the information, including Scopus, Science Direct, Google Scholar, PubMed, and Web of Science. Results and conclusions: Although many of its traditional uses have gradually faded into obscurity over the centuries, recent decades have rekindled the interest in this species. Recent ethnobotanical surveys have reported the use of this species against various health-related conditions, while current pharmacological studies have corroborated several health benefits of the species, such as antioxidant, antidiabetic, anti-hyperpigmentation, anticancer, and antimicrobial activities. The validated properties are mainly attributed to the presence of multiple phytoconstituents belonging to flavonoids, phenolic acids, terpenes, and fatty acids, which could also indicate potential uses for T. balsamita in the food industry as a natural preservative and flavoring agent of food products. Full article

Background/Objectives: The integration of Artificial Intelligence (AI) and Machine Learning (ML) in pharmaceutical research and development is transforming the industry by improving efficiency and effectiveness across drug discovery, development, and healthcare delivery. This review explores the diverse applications of AI and ML, emphasizing their role in predictive modeling, drug repurposing, lead optimization, and clinical trials. Additionally, the review highlights AI’s contributions to regulatory compliance, pharmacovigilance, and personalized medicine while addressing ethical and regulatory considerations. Methods: A comprehensive literature review was conducted to assess the impact of AI and ML in various pharmaceutical domains. Research articles, case studies, and industry reports were analyzed to examine AI-driven advancements in predictive modeling, computational chemistry, clinical trials, drug safety, and supply chain management. Results: AI and ML have demonstrated significant advancements in pharmaceutical research, including improved target identification, accelerated drug discovery through generative models, and enhanced structure-based drug design via molecular docking and QSAR modeling. In clinical trials, AI streamlines patient recruitment, predicts trial outcomes, and enables real-time monitoring. AI-driven predictive maintenance, process optimization, and inventory management have enhanced efficiency in pharmaceutical manufacturing and supply chains. Furthermore, AI has revolutionized personalized medicine by enabling precise treatment strategies through genomic data analysis, biomarker discovery, and AI-driven diagnostics. Conclusions: AI and ML are reshaping pharmaceutical research, offering innovative solutions across drug discovery, regulatory compliance, and patient care. The integration of AI enhances treatment outcomes and operational efficiencies while raising ethical and regulatory challenges that require transparent, accountable applications. Future advancements in AI will rely on collaborative efforts to ensure its responsible implementation, ultimately driving the continued transformation of the pharmaceutical sector. Full article

Background/Objectives: This systematic review explores the utilization of electrospinning for the incorporation of natural compounds, focusing on their pharmacological applications. Methods: This systematic review focused on studies investigating the incorporation of natural bioactive compounds into nanofibers produced via the electrospinning technique for pharmacological applications. The search was conducted for English-language articles published online between 1 January 2013 and 10 December 2023. The review followed a structured approach, excluding review articles, clinical studies, and gray literature such as unpublished works, non-peer-reviewed journals, theses, and industry data. Results: The review of 99 articles highlighted the advantages of electrospun nanofibers in tissue regeneration, infection control, and controlled drug release, with notable potential in oncology for targeted antitumor drug delivery. It discussed the influence of polymers and solvents on fiber characteristics and identified a significant gap in cosmetic applications, emphasizing the technique’s potential for prolonged release and improved ingredient stability. Additionally, this review noted the underutilization of marine-derived substances, which possess rich bioactive properties that could benefit biomedical and cosmetic fields. Conclusions: This systematic review highlights the advantages of electrospinning for pharmacological applications, including tissue regeneration, infection control, and controlled drug release, with promising potential in oncology. However, gaps were identified in the cosmetic field and the use of marine-derived substances. Future advancements in electrospinning technology and interdisciplinary collaboration are essential to unlocking its full potential in medicine and cosmetics. Full article

Nicotinamide adenine dinucleotide (NAD⁺) is one of the most essential coenzymes that is widely distributed in human tissues. However, with the progress of aging, the NAD⁺ level gradually decreases, thus impacting the metabolic dynamics and heightening susceptibility to various pathologies. Increasing NAD⁺ levels are expected to delay aging and improve age-related degenerative diseases. Amino–carboxylic semialdehyde dehydrogenase (ACMSD) is a key enzyme involved in the de novo synthesis of NAD⁺. It reduces the intermediate products of the de novo synthesis pathway by catalyzing the degradation of α-amino-β-carboxyethylglutamic acid-ε-semialdehyde (ACMS), thus reducing the production of NAD⁺. Genetic and pharmacological inhibition of ACMSD has been demonstrated to increase NAD⁺ levels in vitro and in vivo, thus making it a potential target for the treatment of NAD⁺-deficient diseases. In this mini-review, we detail the molecular mechanisms regulated by ACMSD. We also discuss the potential efficacy and progress of ACMSD inhibitors in treating aging and age-related diseases. Full article

Background/Objectives: The marine sponge Plakortis halichondrioides is notable for its capacity to accumulate a large array of secondary metabolites. The present research aims at discovering new secondary metabolites from P. halichondrioides with potential applications in medicine. Methods: Plakortilactone (1) and seco-plakortide F acid (2), two new polyketides, along with known manadodioxan D (3), 13-oxo-plakortide F (4), plakortide F (5), and manadodioxan E (6), were isolated from P. halichondrioides. We achieved the structural elucidation of 1 and 2 using modern spectroscopic methods. The relative stereochemistry of 1 was proposed on the basis of 1D- and 2D-NMR data in combination with molecular modeling studies. Additionally, the absolute configuration of 2 was established through chemical correlation to 5. We screened some of the isolated compounds against the malaria parasite Plasmodium falciparum 3D7 non-resistant (wild-type resistant) strain, the tuberculosis bacterium Mycobacterium tuberculosis, the prostate cancer cell line DU-145, and the melanoma cell line A-2058. Results: minimal activity was detected for 2 against these cancer cells. In contrast, 3 and 4 displayed activity against DU-145 cells with IC₅₀ values of 1.6 µg/mL and 4.5 µg/mL, respectively, and A-2058 cells with IC₅₀s of 2.6 µg/mL and 7.7 µg/mL, respectively. In the antiplasmodial activity assay 3 appeared more active (IC₅₀ = 1.7 µg/mL) than 4 (IC₅₀ = 3.1 µg/mL). Meanwhile, 2 displayed only moderate activity (IC₅₀ = 39.3 µg/mL). In the antimycobacterial activity assay 2 exhibited moderate inhibition (MIC = 75.1 µg/mL). In contrast, a 1:1 mixture of 3 and 4 demonstrated higher activity (MIC = 26.3 µg/mL). Conclusions: the biological activity data together with ADMET predictions indicated favorable pharmacokinetic properties for 3. Full article

COVID-19, first identified in December 2019 in Wuhan, China, is caused by the SARS-CoV-2 virus, a pathogen that primarily targets the respiratory system and can lead to severe conditions such as acute respiratory distress syndrome (ARDS). Among the seven coronaviruses known to infect humans, three—SARS-CoV, MERS-CoV, and SARS-CoV-2—are associated with severe illness and significant morbidity. SARS-CoV-2 is an enveloped, single-stranded RNA virus that utilizes the angiotensin-converting enzyme 2 (ACE2) receptor for cellular entry. The genetic sequence of SARS-CoV-2 is highly mutable, leading to the emergence of variants that alter disease pathology and transmission dynamics. The World Health Organization (WHO) has classified these mutations into variants of concern (VOCs), variants of interest (VOIs), and variants under monitoring (VUMs). This review provides an in-depth analysis of both historical and emerging SARS-CoV-2 variants, summarizes recent advancements in diagnostic methods for SARS-CoV-2 detection, and discusses current therapeutic strategies for COVID-19, with a particular focus on virus-like particle (VLP) vaccines developed in recent years. Additionally, we highlight ongoing therapeutic approaches and their implications for managing COVID-19. Full article

Background/Objectives: The interaction of bioactive compounds derived from plants with drugs has become a significant area of investigation due to its potential to improve, reduce, or have no effect on therapeutic outcomes. Due to the dual effect of these interactions, elucidating the underlying mechanisms is essential for establishing a therapeutic strategy. This study emphasizes the significant findings, mechanisms, and clinical implications of drug–plant bioactive interactions. It calls for more studies to seek safe and effective incorporation into clinical practice. Methods: To identify relevant studies, we performed a systematic literature search based on various scientific databases from 11 August 2024 to 30 December 2024. The search will be based on relevant keywords such as synergy, antagonism, plant bioactive compounds, and drug interactions supplemented with secondary terms such as phytochemicals, herb-drug interactions, pharmacokinetics, and pharmacodynamics. Results: Plant bioactives, including polyphenols, flavonoids, alkaloids, and terpenoids, display valuable biological activities that can interact with medications in three principal ways: synergy, additive effects, and antagonism. Synergy occurs when the combined effects of plant chemicals and pharmaceuticals outweigh the sum of their separate effects, increasing therapeutic effectiveness or allowing dosage decrease to reduce adverse effects. Additive effects occur when the combined impact equals the total individual effects, resulting in better outcomes without increasing risk. Antagonism occurs when a plant ingredient reduces or counteracts the effects of a medicine, thereby jeopardizing treatment. In addition, specific interactions may have no discernible effect. The chemical makeup of bioactive chemicals, medication pharmacokinetics, and individual patient characteristics such as genetics and metabolism all impact the intricacy of these interactions. Conclusions: Pharmacokinetics and pharmacodynamics of drugs can be considerably modulated through their interactions with plant bioactive components, which may cause a significant decrease in efficacy or increase in toxicity of therapeutic agents. More studies are needed to clarify mechanisms of action, prove clinical relevance, and create guidelines for safe co-administration. This integrative approach can mitigate those risks and allow for therapeutic optimization by introducing pharmacogenomics and personalized medicine approaches. Full article

Background/Objectives: Cancer is a leading cause of death. However, recently developed immunotherapies have shown significant promise to improve cancer treatment outcomes and survival rates. Pembrolizumab, a cancer immunotherapy drug, enables a strong T-cell response specifically targeting cancer cells to improve patient outcomes in more than 16 types of cancer. The increasing demand for pembrolizumab, the highest selling drug in 2023, increases global dependence on drug production, which can be vulnerable to supply chain disruptions. Methods: Cell-free protein synthesis (CFPS) is a rapid in vitro protein production method that could provide the production of an immunotherapy drug in an emergency and could facilitate on-demand production of the therapeutic at the point of care if needed. Furthermore, CFPS has potential as a production platform of biosimilars, as the patent for pembrolizumab is set to expire in 2028. Results: This work presents the design, synthesis, and target-binding affinity of a novel single-chain variable fragment of pembrolizumab (Pem-scFv) using CFPS. The CFPS production of Pem-scFv also enables the direct optimization of synthesis reaction composition and expression conditions. The conditions of 30 °C, 35% (v/v) cell extract, and an oxidizing redox environment resulted in the highest Pem-scFv soluble yield of 442 µg/mL. An affinity assay demonstrated significant binding between the CFPS-produced Pem-scFv and the PD-1 target. Computational simulations of Pem-scFv folding and binding corroborate the experimental results. Full article